1 Guidance for Industry

2

3 Drugs, Biologics, and Medical Devices 

4 Derived from Bioengineered Plants for 

5 Use in Humans and Animals

6

7

8 DRAFT GUIDANCE

9

10

11 This guidance document is being distributed for comment purposes only.

12

13

14 Submit comments and suggestions regarding this draft document by the date provided in the 

15 Federal Register notice announcing the availability of the draft guidance. Submit comments to 

16 Dockets Management Branch (HFA-305), Food and Drug Administration, 5600 Fishers Lane, 

17 rm. 1061, Rockville, MD 20852. You should identify all comments with the docket number 

18 listed in the notice of availability that publishes in the Federal Register.

19

20 For questions on the content of this draft document as it relates to FDA-regulated products, 

21 contact Keith Webber, Ph.D., at 301-827-0850 (CBER), Yuan Yuan Chiu, Ph.D., at 301-827-

22 5918 (CDER), Wendelyn Jones Warren, Ph.D., at 301-827-6978 (CVM). For questions 

23 regarding veterinary biological products, contact Patricia L. Foley, D.V.M., Ph.D., at 515-232-

24 5785 (USDA/APHIS/CVB).

25

26 U.S. Department of Health and Human Services

27 Food and Drug Administration

28 Center for Biologics Evaluation and Research (CBER)

29 Center for Drug Evaluation and Research (CDER)

30 Center for Food Safety and Applied Nutrition (CFSAN)

31 Center for Devices and Radiological Health (CDRH)

32 Center for Veterinary Medicine (CVM)

33 U.S. Department of Agriculture

34 Animal and Plant Health Inspection Service (APHIS)

35 Center for Veterinary Biologics (CVB)

36 Biotechnology Regulatory Services (BRS)

37 September 2002
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38 Guidance for Industry

39

40 Drugs, Biologics, and Medical Devices Derived from Bioengineered 

41 Plants for Use in Humans and Animals

42

43

44 Additional copies of this guidance are available from:

45 Office of Communication, Training, and Manufacturers Assistance, HFM-40

46 Center for Biologics Evaluation and Research

47 Food and Drug Administration

48 1401 Rockville Pike, Rockville, MD 20852-1448

49 Phone: 301-827-4573

50 Internet: http://www.fda.gov/cber/guidelines.htm

51 Mail: The Voice Information System at 800-835-4709 or 301-827-1800

52 or

53 Office of Training and Communication

54 Division of Communications Management Drug Information Branch, HFD-210

55 Center for Drug Evaluation and Research

56 Food and Drug Administration

57 5600 Fishers Lane, Rockville, MD 20857

58 Phone: 301-827-4573

59 Internet: http://www.fda.gov/cder/guidance/index.htm

60 or

61 Division of Small Manufacturers, International, and Consumer Assistance (DSMICA), HFZ-220

62 Center for Devices and Radiological Health

63 Food and Drug Administration

64 1350 Piccard Drive, Rockville, MD 20850

65 800-638-2041 or 301-443-6597

66 Internet: http://www.fda.gov/cdrh

67 Email: DSMICA@cdrh.fda.gov

68 Facts-On-Demand (faxback): 301-827-0111

69 or

70 Communications Staff, HFV-12

71 Center for Veterinary Medicine (CVM)

72 Food and Drug Administration

73 7500 Standish Place

74 Rockville, MD 20855

75 Phone: 301-594-1755

76 Internet :http://www.fda.gov/cvm
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77 Table of Contents

78

79 I. INTRODUCTION............................................................................................................. 1

80 A. Purpose and Scope ................................................................................................ 1

81 B. Regulatory Responsibility .................................................................................... 2

82 II. HOST AND SOURCE PLANT CHARACTERIZATION ........................................... 3

83 A. General Considerations ........................................................................................ 3

84 B. Host Plants............................................................................................................. 3

85 C. Bioengineered Source Plants................................................................................ 4

86 III. ENVIRONMENTAL CONSIDERATIONS................................................................... 7

87 A. General Considerations ........................................................................................ 7

88 B. National Environmental Policy Act (NEPA) ...................................................... 8

89 C. Confinement Measures......................................................................................... 8

90 IV. MANUFACTURING AND PROCESS-RELATED CONSIDERATIONS............... 11

91 A. General Considerations ...................................................................................... 11

92 B. Special Considerations for Whole Fruit or Vegetable Products .................... 12

93 C. Applicable FDA and USDA Regulations .......................................................... 12

94 D. Product Manufacturing Procedures................................................................. 13

95 E. Characterization of the Product ........................................................................ 17

96 F. Product Stability ................................................................................................. 17

97 V. PRE-CLINICAL CONSIDERATIONS FOR BIOENGINEERED 

98 PHARMACEUTICAL PLANT-DERIVED PRODUCTS FOR USE IN HUMANS 18

99 A. General Considerations...................................................................................... 18

100 B. Evaluation of Impurities..................................................................................... 19

101 C. Allergenicity......................................................................................................... 20

102 D. Immunogenicity................................................................................................... 20

103 VI. CLINICAL TESTING FOR FDA-REGULATED PRODUCTS AND PRE104 LICENSE TESTING FOR USDA-REGULATED PRODUCTS ............................... 20

105 VII. DEFINITIONS ................................................................................................................ 21

106 VIII. REFERENCES................................................................................................................ 24

107 APPENDIX A.............................................................................................................................. 26

108
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108 Guidance for Industry

109

110 Drugs, Biologics, and Medical Devices Derived from Bioengineered 

111 Plants for Use in Humans and Animals

112

113

114 This guidance document represents the agencies’ current thinking on this topic. It does not 

115 create or confer any rights for or on any person and does not operate to bind FDA, USDA, or 

116 the public. An alternative approach may be used if such approach satisfies the requirements of 

117 applicable statutes and regulations.

118

119

120 I. INTRODUCTION

121

122 A. Purpose and Scope

123

124 This document is the result of a combined effort by the U.S. Food and Drug Administration 

125 (FDA) and the U.S. Department of Agriculture (USDA) to provide guidance with regard to 

126 the use of bioengineered plants or plant materials to produce biological products, including 

127 intermediates, protein drugs, medical devices, new animal drugs, and veterinary biologics 

128 regulated by FDA or USDA (hereafter referred to as “regulated products”). This document 

129 does not address non-protein drugs, botanicals, or allergenic products (21 CFR 680.1) for 

130 human use. It should be noted, however, that if a bioengineered pharmaceutical plant is 

131 used to produce a non-protein drug product, the principles described in this document 

132 regarding the host and source plant characterization and the environmental considerations 

133 would be applicable. If you are planning to produce a non-protein drug product for human 

134 use in a bioengineered pharmaceutical plant, consultation with FDA’s Center for Drug 

135 Evaluation and Research (CDER) early in the drug development process is encouraged. For 

136 the purposes of this document, the term “bioengineered pharmaceutical plant” means any 

137 plant manipulated by recombinant DNA technology to express a gene encoding a biological 

138 or drug product. 

139

140 Within this document, “you” refers collectively to sponsors, manufacturers, licensees, and 

141 applicants; “we” refers to FDA and/or USDA/Animal and Plant Health Inspection Service 

142 (APHIS)/Center for Veterinary Biologics (CVB).

143

144 This document outlines important scientific questions and information that you should 

145 address during the investigation of a new animal drug and preparation of an Investigational 

146 New Drug (IND) application, Investigational Device Exemptions (IDE), Biologic License 

147 Application (BLA), New Drug Application (NDA), New Animal Drug Application 

148 (NADA), Premarket Approval (PMA), or 510(k) to the FDA, or a United States Veterinary 

149 Biological Product License Application (VBPLA) to the USDA (hereafter referred to as 

150 “your application”). This document presents points that you should consider to demonstrate 

151 the safety and effectiveness of products from bioengineered pharmaceutical plants for use in 
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152 humans or animals or as components in clinical diagnostic systems. 

153

154 In addition, this document presents points you should consider in addressing environmental 

155 issues as well as confinement measures that should be an integral part of the manufacturing 

156 process for all pharmaceutical products produced in bioengineered pharmaceutical plants or 

157 plants infected with engineered vectors containing genetic material for the expression of 

158 regulated products. 

159

160 This document is directed at the issues unique to the use of bioengineered pharmaceutical 

161 plants as source material for the production of FDA and/or USDA regulated products. 

162 Therefore, it does not focus on many aspects of regulated products that are shared with other 

163 expression systems. Given the complexity and variety of products, no single document can 

164 anticipate and address all issues. You are encouraged to consult other FDA and USDA 

165 documents for guidance on other specific topics relevant to your product.

166

167 You should be aware that the Biotechnology Regulatory Services Division (BRS) within 

168 APHIS oversees the importation and interstate movement of bioengineered pharmaceutical 

169 plants and infectious plant vectors as well as the release of these entities into the 

170 environment (i.e., outside of a contained facility, such as a greenhouse, laboratory, or 

171 fermentor). You must receive a permit from APHIS/BRS prior to engaging in these 

172 activities (7 CFR 340). You may obtain guidance on applying for a permit at the 

173 USDA/APHIS website http://www.aphis.usda.gov/biotech or by writing to 

174 USDA/APHIS/BRS (see addresses in Appendix A). This document will not describe the 

175 plant permitting process. 

176

177 B. Regulatory Responsibility

178

179 The FDA regulates human biologics, and human and animal drugs derived from 

180 bioengineered pharmaceutical plants, intended for therapeutic, preventative, or diagnostic 

181 purposes. Biological products and drugs for use in humans are regulated by the Center for 

182 Biologics Evaluation and Research (CBER) and CDER under authority of the Public Health 

183 Service Act (PHS Act) (42 U.S.C. 262 et seq.) and the Federal Food, Drug, and Cosmetic 

184 Act (FD&C Act) (21 U.S.C. 301 et seq.). FDA also regulates animal drugs derived from 

185 bioengineered pharmaceutical plants, intended for use in the diagnosis, cure, mitigation, 

186 treatment, or prevention of disease in animals or to alter the structure or function of the 

187 animal. New animal drugs and animal feeds containing new animal drugs are regulated by 

188 the Center for Veterinary Medicine (CVM) under authority of the FD&C Act. The FDA 

189 regulations are found at Title 21 of the Code of Federal Regulations (21 CFR). 

190

191 The USDA regulates veterinary biologics through the Center for Veterinary Biologics 

192 (CVB) within Veterinary Services in APHIS under the authority of the Virus, Serum, and 

193 Toxins Act (21 U.S.C. 151 et seq.). The USDA regulations are found at Title 9 of the Code 

194 of Federal Regulations (9 CFR) Parts 101-124. 

195

196 As mentioned above, APHIS/BRS regulates the importation, interstate movement, and 

197 release into the environment (e.g., field testing) of all such bioengineered pharmaceutical 
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198 plants, under the Plant Protection Act (7 U.S.C. 7701-7772). The APHIS/BRS regulations 

199 are found at Title 7 of the Code of Federal Regulations (7 CFR), in particular 7 CFR 340.

200

201 Appendix A provides a listing of the points of contact at the agencies.

202

203 To minimize duplication, review of environmental safety issues posed by field growth of the 

204 bioengineered pharmaceutical plants, including National Environmental Policy Act (NEPA) 

205 assessments, will be addressed primarily by APHIS/BRS. Because bioengineered 

206 pharmaceutical plants will be grown under APHIS permit, and because permits enabling 

207 field trials will be obtained prior to submission of a product application, APHIS/BRS will 

208 identify and evaluate the potential environmental effects posed by field growth of such 

209 plants. Environmental concerns posed by use of the regulated product will be addressed in 

210 the NEPA analysis conducted by the regulatory agency responsible for review and/or 

211 approval of the product. These agencies' NEPA analyses will take into account 

212 APHIS/BRS’s environmental reviews. Also refer to section III.B. National Environmental 

213 Policy Act.

214

215

216 II. HOST AND SOURCE PLANT CHARACTERIZATION

217

218 A. General Considerations

219

220 In the development stage, you should give careful consideration to choosing the plant 

221 species that will be used as the source of the desired regulated product. Concerns to be 

222 addressed include: the potential for the plant to express an allergenic or toxic compound; the 

223 method of plant propagation and the measures to ensure confinement; and, if it is a food 

224 crop species engineered to produce non-food material, the measures to ensure that non-food 

225 (or non-feed) material will not get into food or feed. The presence of any such material in 

226 food or feed could render such products adulterated under the FD&C Act (21 U.S.C. 342).

227

228 You are encouraged to refer to pertinent guidance documents and regulations, and to consult 

229 with the regulatory agencies as early as possible in the development process to ensure that 

230 you are aware of the most current regulatory requirements. 

231

232 B. Host Plants

233

234 You should provide in your application a thorough description of the host plant biology that 

235 includes information necessary to identify it in the narrowest taxonomic grouping applicable 

236 (e.g., genus, species, subspecies, variety or cultivar, line designation).

237

238 In order for the agencies to assess the ability of the chosen plant to consistently manufacture 

239 your intended product, you should submit a description of the reproductive biology of the 

240 unmodified plant and production practices with regard to:

241 • growth habitat as an annual, perennial, or biennial;

242 • timing of sexual maturity and duration of flowering;

243 • seed production and harvesting;
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244 • recognized practices for maintaining seed stock purity;

245 • conditions of growth;

246 • timing of harvest;

247 • method of harvesting; and

248 • transporting, storage and sorting of harvested materials.

249

250 In addition, you should provide a description of the host plant including levels of any toxins, 

251 anti-nutrients, and allergens known to be produced by the plant species and whether it is 

252 known to accumulate heavy metals. Please state if the plant is of a species used for food or 

253 feed in a raw or processed form. 

254

255 C. Bioengineered Source Plants 

256

257 1. General Considerations

258

259 The host plant may be bioengineered to increase the expression of an endogenous 

260 gene product or to manipulate the plant to produce a heterologous gene product. 

261 The modifying gene may be transiently added to the plant or it may be inserted in a 

262 stable manner. Regardless of the method of gene expression used, traceable 

263 documentation of the growth and expression phase of the manufacturing process, 

264 including banking of the plant lines and/or vectors should be maintained. Most 

265 importantly, you should include data in your application to demonstrate that the 

266 source plant produces a consistent product.

267

268 When the bioengineered pharmaceutical plant is from a species that is used for food 

269 or feed, measures should be in place to ensure that there is no inadvertent mixing of 

270 the bioengineered plant material with plant material intended for food or feed use. 

271 The presence of any such material in food or feed could render such products 

272 adulterated under the FD&C Act (21 U.S.C. 342). We strongly recommend that you 

273 have tests available that can detect the presence of the target gene and the protein 

274 product in the raw agricultural commodity.

275

276 2. Characterization of the Recombinant DNA

277

278 In your application, you should provide a full characterization of the recombinant 

279 DNA constructs or viral vectors used to transfer genes, including:

280 • the origin and function of all component parts of the construct, including 

281 coding regions, antibiotic- or herbicide-resistance genes, origins of replication, 

282 promoters, and enhancers; 

283 • physical map of the construct(s) illustrating the position of each functional 

284 component; 

285 • method used for plasmid propagation;

286 • any sequences required for bacterial expression of plasmid constructs;

287 • the nucleotide sequence of the intended insert up to and including the 

288 junctions at the 5’- and 3’- ends; and 

289 • any changes in codons to reflect more acceptable codon usage in plants.
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290

291 For the purposes of this document, coding regions include full-length and

292 truncated sense constructs, antisense constructs, and constructs containing 

293 ribozymes, regardless of whether or not the coding region is designed or expected 

294 to be expressed in the bioengineered pharmaceutical plant.

295

296 For additional details regarding analysis of r-DNA constructs for human 

297 biologics, please refer to the International Conference on Harmonisation (ICH); 

298 Technical Requirements for Registration of Pharmaceuticals for Human Use –

299 Guideline Q5B: Quality of Biotechnological Products: Analysis of the Expression 

300 Construct in Cells Used for Production of r-DNA Derived Protein Products (Ref. 

301 1). 

302

303 3. Stable Transformation Systems

304

305 Before preparing Master Seeds or Master Seed Banks (MSB) and Working Seeds 

306 or Working Seed Banks (WSB), we recommend that you establish a suitable 

307 transformant. For stable transformation systems, you should describe the gene 

308 transfer method in detail and provide relevant references, as appropriate. An 

309 analysis should be performed to determine the number of copies of the gene 

310 inserted, the number of integration sites, and to demonstrate if complete or partial 

311 copies are inserted into the plant’s genome. You should determine the nucleotide 

312 sequence of the insert from DNA or mRNA retrieved from the stably-transfected 

313 plants in order to confirm the integrity and fidelity of the DNA insert. When a 

314 fragment of a coding region designed to be expressed in a plant is detected, you 

315 should determine whether a fusion protein could be produced and in which host 

316 tissues it may be located. 

317

318 If the transformation system utilizes a pathogenic organism or nucleic acid 

319 sequences from a pathogen, you should provide a description of the pathogen, the 

320 strain, and the gene(s) involved. If any such pathogenesis-related DNA sequences 

321 were removed or altered prior to transformation, you should describe these 

322 changes in detail. Any helper plasmids or analogous DNA fragments used in the 

323 transformation process should also be described. For example, for 

324 Agrobacterium-mediated transformation, provide the strain designation of the 

325 Agrobacterium used during the transformation process, indicate how the Ti 

326 plasmid-based vector was disarmed, and indicate whether Agrobacterium was 

327 cleared from the transformed tissue.

328

329 You should submit a complete description of the process, including selection 

330 methods for the final transformant. You should include the source of and the 

331 methods used to prepare the recipient tissue or cells and, if the tissues or cells are 

332 cultured or pre-treated in any way, you should provide a complete description of 

333 the reagents used and composition of the culture medium. For direct 

334 transformation methods, you also should provide a thorough description of the 

335 transforming DNA preparation: including amount and concentration of transgenic 
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336 DNA; the nature, source, and concentration of any carrier DNA; the composition 

337 and source of carrier particles; and the source and concentration of any other 

338 excipients. In addition, you should describe in detail any tests used to evaluate 

339 the transformations process and provide the results.

340

341 4. Transient Transfection Systems:

342

343 Virus-mediated transient transfection systems, in their simplest form, employ two 

344 components: a recombinant virus vector and a host plant. Characterization of the 

345 host plant should include the information outlined in section II. B., above. The 

346 information you provide regarding the recombinant virus vector should include 

347 the following: 

348 • the taxonomic name of the virus, including family, genus, and strain 

349 designation, including any synonyms;

350 • the type of nucleic acid contained in the virus (DNA or RNA); 

351 • whether the virus is associated with any satellite or helper viruses;

352 • the natural host range of the virus; 

353 • how the virus is transmitted;

354 • if the virus is transmitted by a vector, the identity of the vector including 

355 mode of transmission (e.g., persistent or non-persistent);

356 • the identity of the viral gene(s) (if known) involved in vector transmission; 

357 • whether any synergistic or transcapsidation interactions with other viruses 

358 under field situations have been reported in the literature;

359 • the protocol for purification of the virus;

360 • the protocol for cloning of recombinant virus; 

361 • a description of the preparation of the Master Plasmid Bank (MPB), if one is 

362 used; 

363 • the storage conditions and data demonstrating stability of the MPB;

364 • the protocol for the preparation of infectious nucleic acid from plasmid; and

365 • data characterizing the infectious nucleic acid with respect to its identity with 

366 the parental genome.

367

368 You should include relevant literature citations to any of the above information, 

369 as appropriate. 

370

371 5. Genetic Stability: Seed Banks and Vegetative Propagation

372

373 Regardless of whether a transient-transfection system or a stable transformation 

374 system is used, you should prepare a MSB and a WSB to ensure consistent lot-to375 lot growth of the plant and expression of the regulated product. The description 

376 of the MSB in your application should include the identification, the method of 

377 production, the results of analytical tests used to characterize it, the size of the 

378 bank, the storage conditions, and data demonstrating its viability, bioburden 

379 (including speciation of contaminants), uniformity of gene content, and stability.

380
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381 You should submit data demonstrating that bioengineered pharmaceutical plant 

382 lines derived through stable transformation are stable in both phenotype and 

383 genotype. To demonstrate genetic stability, you should include data from a 

384 segregation analysis for the trait of interest, as well as from a molecular 

385 characterization of the genomic insert (e.g., Southern analysis) and from analyses 

386 of expression of the intended product.

387

388 For plants that are fertile, you should provide data demonstrating the pattern and 

389 stability of inheritance and expression of the new traits over several generations

390 sufficient to ensure stability over the number of generations that will be used 

391 during manufacture of the regulated product. 

392

393 For plants that are infertile or for which it is difficult to produce seed (such as 

394 vegetatively propagated male-sterile potatoes), you should provide data to 

395 demonstrate that the trait is stably maintained and expressed during vegetative 

396 propagation over a number of cycles that is appropriate to the crop.

397

398 6. Tissue Distribution of Expression Products

399

400 For all inserted coding regions, you should provide data that demonstrates 

401 whether the protein is or is not produced (describe assay method and indicate 

402 limit of detection) as intended in the expected tissues consistent with the 

403 associated regulatory sequences driving its expression (e.g., if the gene is 

404 inducible, you should determine if the gene is expressed in the expected tissues 

405 under induction conditions). You should provide quantitative data characterizing 

406 the distribution of the product in the major plant tissues (e.g., leaves, roots, stalks, 

407 seeds).

408

409

410 III. ENVIRONMENTAL CONSIDERATIONS

411

412 A. General Considerations

413

414 Using bioengineered pharmaceutical plants to produce regulated products for use in 

415 animals or humans raises a number of environmental concerns that you should address, 

416 including confinement measures that may be needed to control the spread of the 

417 bioengineered pharmaceutical plants and to keep them from entering the food or feed 

418 supply. We encourage you to consult with the regulatory agencies as early as possible in 

419 the development process to ensure that you are aware of the most current regulatory 

420 requirements. For example, you should contact APHIS/BRS for more information on 

421 regulations governing the plants while in the field or in transport. APHIS/BRS 

422 authorization is required for the interstate movement, importation, and field release of 

423 plants addressed by this guidance (7 CFR 340). For most initial experiments and 

424 commercial uses of these plants, a USDA/APHIS/BRS permit will be needed. Refer to 

425 USDA regulations (7 CFR 340) that can be found at APHIS's home page 

426 http://www.aphis.usda.gov/biotech. 
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427

428 Bioengineered pharmaceutical plants that are grown exclusively in an enclosed building 

429 (e.g., greenhouse) generally will be considered to be confined during the growing period 

430 if there are control measures in place to eliminate the spread of pollen or seeds outside of 

431 the facility. Growing plants in such an enclosed building does not require a 

432 USDA/APHIS/BRS permit, however, the importation or interstate movement of

433 bioengineered pharmaceutical plants would require a permit (7 CFR 340.4).

434

435 B. National Environmental Policy Act (NEPA)

436

437 You should be aware of NEPA requirements for both the FDA (21 CFR part 25) and the 

438 USDA (7 CFR part 372). You should consider the potential environmental impact of all 

439 aspects of the manufacturing process, including but not limited to transport of seeds and 

440 plants, planting, growing, harvesting, processing, purifying, packaging, storage, and 

441 disposal. If you believe that your activities are categorically excluded by 7 CFR 

442 372.5(c), 21 CFR 25.31, or 25.33 from the requirement to submit an environmental 

443 assessment, you should state this in your application. You are encouraged to consult 

444 available guidance documents (Refs. 2, 3) and to talk directly with the USDA and the 

445 FDA regarding NEPA requirements. A copy of the letter from APHIS/BRS granting 

446 your permit should be submitted in your application for the regulated product in support 

447 of the environmental assessment (21 CFR 25.15 and 25.40) or the claim of categorical 

448 exclusion (21 CFR 25.31, 25.33 or 7 CFR 372.5(c)). FDA and CVB intend to take 

449 APHIS/BRS evaluations and determinations into account in doing their own NEPA 

450 assessments.

451

452 C. Confinement Measures

453

454 1. General Considerations

455

456 Regardless of whether the bioengineered pharmaceutical plants are grown and/or 

457 processed by you or on a contractual basis by other persons, manufacturing 

458 controls are your responsibility and should be documented clearly in standard 

459 operating procedures (SOPs), Outlines of Production, or other records, as 

460 appropriate (see section IV.C., Applicable FDA and USDA Regulations). For 

461 FDA regulated products, refer to 21 CFR 200.10, parts 210 and 211, 514.1, and 

462 820.50; see also FDA’s Draft Guidance for Industry: Cooperative Manufacturing 

463 Arrangements for Licensed Biologics (Ref. 4) once it is finalized. 

464

465 In developing a bioengineered pharmaceutical plant, you should implement 

466 procedures to ensure that such a plant line is used only for its intended purpose as 

467 a source material for a regulated product. As described in 7 CFR 340.4, 340.7, 

468 and 340.8, a permit from USDA/APHIS/BRS is required for the interstate 

469 transport of bioengineered pharmaceutical plants or seeds for such plants, and you 

470 must keep records documenting the handling and transfer of such materials. 

471 Shipment of bioengineered pharmaceutical plants for veterinary biologics requires 

472 permission from USDA/APHIS/BRS. When manufacturing firms are shipping 
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473 veterinary biological products at any stage of production, shipment must be 

474 authorized by CVB and is regulated under 9 CFR 103.3. Such controlled transfer 

475 of source materials helps ensure that these plants are not diverted to unintended 

476 uses. 

477

478 When a plant species that is used for food or feed is bioengineered to produce a 

479 regulated product, you should consider the use of strategies that allow the 

480 bioengineered pharmaceutical plant line to be readily distinguished from its food 

481 or feed counterpart. Such strategies might include the use of genetic markers that

482 alter the physical appearance of the plant (e.g., a novel color or leaf pattern), or 

483 change the conditions under which a plant will grow (e.g., the use of an 

484 auxotrophic marker gene). You should also consider strategies to reduce the 

485 likelihood of unintended exposure to a regulated product by restricting the 

486 expression of the bioengineered pharmaceutical product to a few specific plant 

487 tissues (e.g., the use of tissue specific promoters) or by restricting the conditions 

488 under which the product will be expressed (e.g., use of an inducible promoter). 

489 For such plants that outcross, you may want to consider growing them in regions 

490 of the country where little or none of its food/feed counterparts are grown.

491

492 Measures should be in place to ensure that there is no inadvertent mixing of the 

493 bioengineered pharmaceutical plant with plant material intended for food or feed 

494 (including inadvertent mixing with seeds for food or feed crops). During the 

495 development of your overall production process (from the farm through the final 

496 product), you should determine where in the process inadvertent mixing could 

497 occur and establish appropriate control measures. We strongly recommend that 

498 you have tests available that can detect the presence of the target gene and the 

499 protein product in the raw agricultural commodity. The presence of the target 

500 gene or gene product in food or feed could render such products adulterated under 

501 the FD&C Act (21 U.S.C. 342). You may wish to consult with FDA’s Center for 

502 Food Safety and Applied Nutrition (CFSAN) or with CVM about the legal 

503 implications of any such material getting into food or feed.

504

505 2. Control of Seed Stocks

506

507 You should maintain careful control over the inventory and disposition of viable 

508 seeds to preclude the possibility that such seeds will be used to produce material 

509 that could be used for food or feed production. When seed stocks are produced, 

510 there should be an accounting of the total yield of seed (e.g., by weight or by 

511 volume). Seed stocks should be stored in aliquots of appropriate volume to allow 

512 reasonably accurate accounting of use and disposition. A record of the amount 

513 and disposition of any withdrawals from the seed bank should be made (7 CFR 

514 340.4(b)(12)). Seed stocks should be prominently labeled in accordance with the 

515 permit issued by APHIS/BRS for field growth or interstate shipment of 

516 bioengineered seeds (7 CFR 340.7).

517
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518 3. Field-grown Plants

519

520 You must have a permit from APHIS/BRS to grow bioengineered pharmaceutical 

521 plants in the field (7 CFR 340.4) and must have control over the growing process 

522 from planting through harvesting and over the disposition of remaining crops 

523 and/or crop residue and, if required, over the subsequent use of the field if for 

524 growth of food or feed or as a pasture during subsequent seasons. All persons 

525 involved in field growth of the product should be adequately trained to perform 

526 the duties for which they are responsible. Control measures should include an 

527 accounting of seed that is transferred from seed bank storage to the field for 

528 planting, or for archiving. Documentation of the size and location of all sites 

529 where the bioengineered pharmaceutical plants will be grown, of the control of 

530 pollen spread, and of the subsequent use of the field and destruction of volunteer 

531 plants in subsequent growing seasons should be maintained and provided to the 

532 FDA and CVB, as appropriate. Fields should be unambiguously identified, such 

533 as by Global Position Satellite (GPS) markers. We recommend that you consider 

534 the use of perimeter fencing to help exclude wildlife and escaped livestock. All 

535 fields used to grow source bioengineered pharmaceutical plants are subject to 

536 inspection by the USDA (7 CFR 340.4; 9 CFR 101-108) and/or by the FDA (42 

537 U.S.C. 262; 21 U.S.C. 374). 

538

539 4. Control of Harvested Material

540

541 APHIS requires that appropriate confinement procedures be in place for transport 

542 of the source material from the field or greenhouse to the production facility (7 

543 CFR 340.4(b)(10-12)). During transport, containers of harvested material should 

544 carry a label that clearly indicates that the material, including but not limited to 

545 seeds, leaves, roots, and stems, is not to be used for food or feed or for any 

546 purposes in which residual materials could be used for food or feed (such as 

547 ethanol production). Reconciliation of the quantities of material leaving the 

548 growing facility and arriving at the processing facility should be made. In 

549 manufacturing of a regulated product, records must be kept to document control 

550 over harvested material in accordance with 21 CFR part 211 subpart J, 21 CFR 

551 part 226 subpart E, 7 CFR 340.4, or 9 CFR part 116 and made available for 

552 inspection by the FDA or CVB, as appropriate.

553

554 5. Control at Processing Facilities

555

556 As stated in section III.C.1., you should implement appropriate procedures to 

557 ensure that bioengineered pharmaceutical plants or plant materials do not 

558 unintentionally mix with other plant products, particularly those used as food or 

559 feed. Source plant materials should not be processed in facilities that also are 

560 used for the production of food or feed, such as grain mills, without prior 

561 consultation with USDA/APHIS/BRS and FDA.

562
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563 6. Control of Waste Material

564

565 In-process wastes (e.g., column wash solutions, diafiltration solutions, etc.), 

566 rejected in-process material, and residual source plant material from the 

567 purification process should be treated to inactivate the regulated product prior to 

568 disposal, as appropriate. They should be disposed in a manner to ensure that the 

569 material will not enter the human or animal food chain unless you have 

570 specifically consulted with FDA for the use of this material in food or feed 

571 products. Disposal should conform to local and state regulations. Waste material 

572 from the manufacture of human drug and biological products, or animal drugs 

573 should be disposed of in a safe and sanitary manner (21 CFR 211.50). Veterinary 

574 biologic materials should be disposed of in a manner consistent with 9 CFR 

575 114.15, Disposal of Unsatisfactory Products and By-products, following 

576 Veterinary Services Memorandum 800.56. If, rather than disposal, the residual 

577 material is to be used for a secondary purpose other than a food or feed product, 

578 there should be clear procedures in place to verify the disposition of this material 

579 and by-products and to document that it will not be used for food or feed.

580

581

582 IV. MANUFACTURING AND PROCESS-RELATED CONSIDERATIONS

583

584 A. General Considerations

585

586 Facilities and procedures used for the manufacturing of regulated products should be 

587 designed to prevent contamination and cross-contamination during harvest and processing 

588 of source material. The flow of personnel, material, product, and waste into and out of the 

589 facility should be designed to prevent contamination of the product. You should establish 

590 written procedures for appropriate cleaning, maintenance, and sanitization of equipment and 

591 utensils to prevent malfunctions or contamination that would alter the safety, identity, 

592 strength, quality, or purity of the drug products beyond established requirements (21 CFR 

593 211.67). In controlled areas with specified air classifications, a program for monitoring the 

594 environment for viable and non-viable particulates should be established based on the 

595 criticality of the manufacturing process involved and should include active monitoring of 

596 critical manufacturing processes as they are performed. For FDA-regulated products, 

597 manufacturing controls, including process validation, should be appropriate for the type of 

598 product and stage of development. The regulations governing facilities requirements are 

599 listed in section IV.C., Applicable FDA and USDA Regulations. 

600

601 Because microbiological contaminants can have an adverse effect on product safety, quality, 

602 and stability, we recommend that you establish processing steps to decrease bioburden 

603 levels as the material moves through the manufacturing process (21 CFR 211.80(b)). The 

604 validation activities described in this section should be phased in during the investigational 

605 phase, as the clinical studies progress toward submission of a regulated product application. 

606 It should be noted however that assurance of sterility or limits on bioburden in the final 

607 product may be required as appropriate, depending on the final form and intended use of the 

608 product (e.g., parenteral vs. whole fruit or vegetable). (21 CFR 211.80, 211.100-103, and 
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609 211.113; 21 CFR parts 226, 514, 610, and 820; and 9 CFR part 113.)

610

611 You should only use source materials with appropriate quality attributes for manufacture of 

612 the product. Each lot of source material should be assessed for the presence of foreign 

613 matter. Care should be taken to minimize contaminants (e.g., molds and other agents that 

614 may be present in the source material) that could lead to the inadvertent exposure of 

615 recipients of regulated products to undesirable impurities or could affect product quality 

616 (e.g., microbial proteases).

617

618 For veterinary biologics, manufacturing must be in accordance with an Outline of 

619 Production filed with CVB as required by 9 CFR 114.8 and 114.9. For all other regulated 

620 products, you must document the manufacturing procedure and lot-specific data (21 CFR 

621 part 211 subpart F, 226.102, part 514, and 820.184). You should ensure that source material 

622 is propagated, harvested, and processed in accordance with written standard operating 

623 procedures that will ensure the adequate processing of the plant derived material and specify 

624 the acceptable limits and kinds of contaminants that may be present. Specifications should 

625 be established regarding the health status of the plants at the time of viral infection and/or 

626 harvest. 

627

628 B. Special Considerations for Whole Fruit or Vegetable Products

629

630 One of the challenges in the use of whole vegetables and/or fruits as the delivery system for 

631 edible biologics is the demonstration of batch uniformity and consistency of dose. A 

632 homogenization step to produce a uniform bulk drug substance, such as a puree, juice, or 

633 milled grain may be necessary. Testing could then be conducted on this homogenized 

634 product to demonstrate potency. In addition, if the plant line used for production is known 

635 to be allergenic, you should consult with FDA or CVB, as appropriate, to discuss the safety 

636 and regulatory issues.

637

638 • Packaging for regulated products must comply with applicable regulations. For 

639 FDA-regulated products, packaging should be consistent with 21 CFR parts 210, 

640 211, 226, 314, 514, 600, 610, and 820. Packaging for APHIS/CVB-regulated 

641 products should comply with 9 CFR part 112. Although edible products for 

642 pharmaceutical use in humans, such as whole fruit or vegetable vaccines, are 

643 regulated as biologics, not foods, we generally recommend that you package your 

644 edible biological products in material that conforms to food packaging regulations 

645 (21 CFR 174.5). The plant source must be clearly identified in the label or 

646 packaging material for biologics for use in humans (21 CFR 610.61(p)) or animals 

647 (9 CFR 112). The plant source should be clearly identified in the labeling of both 

648 oral and non-oral prescription drugs (21 CFR 201.57(a)(2); see also 21 CFR 

649 201.100(b)(4) and (5)). For products containing viable seeds, you should consult 

650 with FDA or CVB, as appropriate. 

651

652 C. Applicable FDA and USDA Regulations

653

654 The specific regulations applicable to the manufacture of a regulated product derived from 
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655 bioengineered pharmaceutical plants are based on: the intended recipient of the product (i.e., 

656 human or animal); the intended use of the product (e.g., biologic, drug, or device); and the 

657 intended route of administration (e.g., parenteral vs. oral). The Table below includes, but is 

658 not limited to, the following applicable regulations for specific classes of regulated products 

659 for use in humans or animals. 

660

Planned use Applicable regulations

Human drug or biologic for parenteral 

administration

7 CFR part 340, 21 CFR parts 210, 211, 

312, 314, 600, 601, 610

Human drug or biologic for oral 

administration

7 CFR part 340, 21 CFR 174.5, parts 210, 

211, 312, 314, 600, 601, 610

Biologic device for human use 7 CFR part 340, 21 CFR parts 600, 601, 

610, 812, 814, 820

Animal drug: Type A medicated articles 

and Type B and C medicated feed

7 CFR part 340, 21 CFR parts 225, 226, 

500, 510, 511, 514, 515, 558

Animal drug 7 CFR part 340, 21 CFR parts 210, 211, 

500, 510, 511, 514

Veterinary biologic 7 CFR part 340, 9 CFR parts 101-118

661

662 We encourage you to refer to FDA and CVB guidance documents for additional information 

663 and recommendations specific to the product class. Any exceptions to the regulatory 

664 requirements must be obtained as provided by regulation. For example, the general safety, 

665 sterility, and mycoplasma tests prescribed in 21 CFR 610.11-12 and 610.30 (for biologics 

666 for use in humans) or 9 CFR 113.26-28 (for veterinary biologics) may be inappropriate for 

667 some products (e.g., edible plant material intended for use as an oral dosage form) and 

668 modifications or alternative, but equivalent, methods of demonstrating a product's safety and 

669 sterility may be permitted in accordance with 21 CFR 610.9 or the product may be 

670 exempted in accordance with 9 CFR 113.4 (see Table, above). 

671

672 D. Product Manufacturing Procedures

673

674 1. General Considerations

675

676 Your application should include a description of each step of the purification 

677 process including analytical tests to demonstrate identity, purity, and 

678 concentration, and the levels of product related and non-product related 

679 impurities. This is particularly important if the impurities are determined to be 

680 toxins, allergens, teratogens, or carcinogens. For each process that is not intended 

681 to be sterile, you should describe the procedures to be followed to control 

682 extraneous bioburden and the in-process testing used to monitor the level of 

683 bioburden (see, 21 CFR 211.113, 226.102, 312.23, 314.50(d), 514.1(b), 820.70, 

684 820.181, and 820.184). A summary of the manufacturing, including propagation 

685 of the source material, should be available at the site where the manufacturing 

686 occurred (21 CFR 211 subpart J). You should consult with the appropriate 

687 agency regarding the applicability of these considerations to device components.

688
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689 2. Growth Conditions

690

691 The Chemistry, Manufacturing, and Controls (CMC) section or the Outline of 

692 Production should include information regarding the location of source plant 

693 propagation. For greenhouse-grown material, you should include in the 

694 description the types of containers, the soil mix composition and qualification 

695 criteria, and the greenhouse growth conditions. For field grown material, the 

696 description should include the previous uses of the land (e.g., agricultural and/or 

697 industrial use). We recommend that you establish specification/acceptance 

698 criteria/limits for the soil composition and potential soil contaminants that may 

699 affect the source material. In addition, you should describe the agricultural 

700 methods utilized during crop growth, including specifications regarding the us e of 

701 chemicals and limits on specific agricultural practices (e.g., the use of specified 

702 fertilizers, pesticides, or herbicides, and irrigation practices relative to a specified 

703 harvest time frame, etc.). You should provide in your application a list of 

704 expected pests that will require control during the growth of the bioengineered 

705 pharmaceutical plants. All pest-control measures implemented should be in 

706 accordance with good agricultural practices for the growth of food crops in the 

707 United States. The Pesticide Product Information System (Ref. 5) contains 

708 information concerning all pesticide products registered in the United States. In 

709 order to evaluate the purity of the product, all pest-control interventions should be 

710 described in appropriate SOPs and should be documented in the Batch Record 

711 (for FDA-regulated products) or Outline of Production (for veterinary biologics). 

712 We recommend that you follow current Good Agricultural Practices (e.g., Ref. 6). 

713 If product expression is induced, either chemically or physically, you should 

714 establish criteria to ensure that induction is performed consistently from batch to 

715 batch. (See generally, 21 CFR parts 210, 211, 226, 312, 314, 514, 601, 610, and 

716 820; see e.g., 21 CFR 211.84, 211.186, 312.23(a)(7), 314.50(d), 514.1, 820.50, 

717 and 9 CFR parts 101-118.)

718

719 3. Harvest 

720

721 You should describe the method of harvesting the source material in written 

722 procedures and document the process in production records. You should have 

723 procedures for determining when the harvest will occur in order to ensure lot-to724 lot consistency of the source material. You should establish specifications for the 

725 harvested material with regard to the levels of active component, process-derived 

726 contaminants, significant endogenous impurities, and adventitious agents. For 

727 example, you should describe agricultural practices and training of harvesting 

728 personnel regarding plant source material quality (e.g., assessment of the disease 

729 status of plant for manual harvesting operations, etc.) (21 CFR part 211 subpart 

730 B). You should have written procedures for establishing the necessary training of 

731 personnel engaged in harvesting plants to ensure the quality of the harvested 

732 material (21 CFR 211.25). We recommend the use of dedicated equipment. We 

733 recommend that equipment-cleaning procedures be developed and that cleaning 

734 agents used on harvesting equipment be described (21 FR 211.67). In addition, 
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735 you should consider measures to prevent the contamination of the harvested 

736 source material with equipment lubricants during processing. (21 CFR part 211 

737 subparts F and J; 21 CFR part 226; 21 CFR 314.50(d)(1), 514.(b)(5); 21 CFR part 

738 814 subpart B; 21 CFR 820.70, 820.75, 820.181, 820.250; and 9 CFR parts 101-

739 118). 

740

741 The description of the harvesting process in the CMC section or Outline of 

742 Production should include specifications regarding acceptable conditions of the 

743 plants and a listing of equipment used to harvest the source material, including 

744 power equipment, hand tools, and transport equipment (see Table, above, for 

745 applicable regulations and refer to applicable FDA and CVB guidance 

746 documents). If the equipment is not dedicated to harvesting only the source 

747 material, other uses should be documented. 

748

749 4. Transfer and Storage Conditions

750

751 Of special concern is the transfer of source material from the field or greenhouse 

752 to the manufacturing facility (see section III.C.1., Confinement Measures; for 

753 authorities concerning the movement of plant materials). The source material 

754 should be transported in such a way as to exclude introduction of insects, vermin, 

755 or potential surface contaminants, which may be carried from the farm field or 

756 greenhouse environment, and to ensure that plant material remains confined 

757 within the container during transport. We recommend that during transport, 

758 containers of regulated product material should carry a label that clearly indicates 

759 that the material is not to be used for food or feed.

760

761 If the harvested source material is to be stored prior to further processing, the 

762 storage conditions (e.g., temperature, humidity, volume, density, storage time, 

763 etc.) should be fully described in your application. The material to be stored 

764 should be characterized and all properties that may be reasonably expected to 

765 affect product quality should be identified and appropriate controls should be 

766 specified (e.g., stability of the product, ability to support growth of 

767 microorganisms, residual soil content, presence of foreign material, insects, 

768 vermin). Source material should be stored under appropriate conditions to ensure 

769 that decomposition processes do not increase the concentration of contaminants 

770 above specified levels or adversely affect the desired active pharmaceutical 

771 ingredient. (21 CFR parts 211, 226, 314, 514, 601, and 820, and 9 CFR parts 

772 101-118).

773

774 5. Initial Processing of Source Material 

775

776 Procedures used to process harvested material should be validated. Harvested 

777 material may be processed to lower bioburden or viability, improve its handling 

778 characteristics, bulk consistency, and/or its extractability using various 

779 procedures, including washing, sanitizing, milling of grain, shredding of leaves, 

780 and homogenization of source plant material, fruits or vegetables. The material 
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781 produced by these processes may be intended for further processing or for use as 

782 the final product (e.g., as an oral vaccine). (21 CFR 211.110, 211.186, 226.40, 

783 and 820.75, and 9 CFR parts 101-118).

784

785 6. Extraction

786

787 The extraction process should be designed to efficiently concentrate the active 

788 component or separate it from the rest of the plant material. As with any 

789 purification procedure, the extraction method should not introduce contaminants 

790 into the process intermediate. Acceptance criteria for relevant parameters (e.g., 

791 product concentration, total protein concentration) should be established in order 

792 to verify lot-to-lot consistency. If the drug or biologic is extracted into a soluble 

793 form, it is advisable to implement sterilizing filtration procedures early in the 

794 process. (See generally, 21 CFR 226.40, 312.23(a)(7), 314.50, 514.1(b)(5)(iv), 

795 820.75 and 9 CFR parts 101-118.)

796

797 7. Aseptic Processing

798

799 For those products for which sterility is required, sterility of protein products is 

800 usually achieved through the use of appropriately validated filtration steps. 

801 However, for products for which sterile filtration is not feasible, we recommend 

802 that you use a validated aseptic process. For FDA-regulated products, refer to 21 

803 CFR 211.113, 610.12(g)(4), and 820.75, and current guidance, such as the 

804 Guideline on Sterile Drug Products Produced by Aseptic Processing (Ref. 7) and 

805 Guidance for Industry: For the Submission of Documentation for Sterilization 

806 Process Validation in Applications for Human and Veterinary Drug Products 

807 (Ref. 8). For veterinary biologics, refer to 9 CFR 113.26 and 113.28 for further 

808 information.

809

810 8. Changeover Procedures

811

812 Changeover procedures designed to prevent contamination between harvests of 

813 source material should be in place and documented (21 CFR 211.67, 226.30, and 

814 820.75). These procedures should include clearance of all materials and waste 

815 from the receiving area and plant material processing equipment, and 

816 cleaning/sanitization of surfaces. Pieces of equipment used for harvesting (e.g., 

817 scythe bars, harvested material transportation vehicles) and initial source material 

818 processing (e.g., maceration equipment) are of particular concern in terms of 

819 cross-contamination. We recommend that only one lot of source material be 

820 processed at a time. If multiple lots of source material are to be processed at one 

821 time, segregation procedures should be developed and implemented. Integrity of 

822 processing equipment should be demonstrated or closed systems employed, when 

823 possible. Product contact equipment should be sufficiently cleaned between each 

824 lot operation to prevent product carry-over contamination of subsequent lots.

825
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826 9. Process Validation

827

828 All processes used to manufacture the product should be validated prior to 

829 marketing the regulated product. Laboratory studies may help to establish 

830 appropriate operating and process parameters and may be used in support of the 

831 formal validation study. You should include information and data from validation 

832 protocols and executed validation studies in your application. (21 CFR 211.110, 

833 211.165, 211.194(a)(2), and 226.40)

834

835 E. Characterization of the Product

836

837 You should provide a complete characterization of the regulated product. For purified drug 

838 substances and drug products provide a characterization sufficient to ensure its identity, 

839 strength, quality, and purity (21 CFR 211.160-165, 211.186, 226.58, 312.23(a)(7), 

840 314.50(d)(1)(i), 601.2(a), 820.60, 820.70, 820.75, 820.80-86, and 820.181). You should 

841 include both physicochemical as well as functional assessments. For purified protein 

842 products, the physicochemical description should also include molecular weight, subunit 

843 composition, isoelectric point, post-translational modifications, impurity profile, and other 

844 relevant parameters. Functional assays should evaluate clinically relevant activities of the 

845 product. You should provide a description of the potency assay for the active component. 

846 You should submit information on the sensitivity, specificity, and variability of all assays, 

847 including the data from the material used to prepare clinical/pre-clinical lots and prelicense 

848 serials that were used to set the acceptance limits for the assay.

849

850 In your application, you should provide specifications for the product, including identity, 

851 purity, potency, physicochemical measurements, and measures of stability (21 CFR 

852 211.160(b) or 9 CFR 114.9). If test results are reported for final release of the product, you 

853 should establish estimates of variability and upper and lower limits for each specification. If 

854 the purified drug substance is held prior to further processing, a description of the storage 

855 conditions and verification of its stability under the conditions described should be included 

856 (see section V.). For FDA-regulated biological products, you are encouraged to consult 

857 related guidance documents for general product characterization guidance (Refs. 9, 10). For 

858 new animal drugs, consult with CVM and for veterinary biologics, CVB.

859

860 You should give special consideration to the characterization of edible plant biologics as 

861 noted above (section IV.B.) especially for measurements of identity of the active drug or 

862 biologic, bioburden limits, dose considerations and final presentation of the product (e.g., 

863 juice, puree, whole fruit, etc.).

864

865 F. Product Stability

866

867 Your application should include a stability protocol containing, but not limited to, testing 

868 for:

869 • potency;

870 • physicochemical measurements that are stability-indicating;

871 • moisture, if lyophilized;
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872 • pH, if appropriate;

873 • sterility or control of bioburden;

874 • pyrogenicity, if applicable; and

875 • general safety, if applicable.

876

877 For products intended for use in humans and for new animal drugs, you should submit 

878 information on the stability of the final product and any in-process material at each holding 

879 step (21 CFR 211.166, 226.58(d), 312.23(a)(7)(iv), 314.50(d)(1), 601.2(a), and 820.75). 

880 Additional information for human drugs and biologics can be found in ICH and FDA 

881 guidance documents (Refs. 9, 10), in 21 CFR part 514, and a CVM specific guidance 

882 document for new animal drugs (Ref. 11). FDA has also published a draft guidance 

883 document issued for public comment and an ICH document on human drug and biological 

884 product stability (Refs. 12, 13). For veterinary biologics, you should establish the stability 

885 of the product prior to licensure.

886

887 You should propose an expiration dating period for the final product and designate the 

888 recommended storage conditions. Also, you should define the procedure for determining 

889 the date from which the expiration dating period begins. 

890

891 A plan for an ongoing stability program should be provided in your application. This should 

892 include the protocol to be used, number of final lots/serials to be entered into the stability 

893 protocol each year, and how such lots/serials will be selected.

894

895

896 V. PRE-CLINICAL CONSIDERATIONS FOR BIOENGINEERED 

897 PHARMACEUTICAL PLANT-DERIVED PRODUCTS FOR USE IN HUMANS 

898

899 A. General Considerations

900

901 This section does not attempt to delineate acceptable practices or testing procedures for each 

902 specific technology or particular class of products, but rather is to provide a general 

903 approach to pre-clinical testing of bioengineered pharmaceutical plant-derived products for 

904 use in humans. You should consult with the appropriate reviewing division of the 

905 appropriate agency for pre-clinical requirements for a specific product class.

906

907 The extent of pre-clinical testing will be determined by the known attributes of the product, 

908 the donor genetic material, the host plant, and the extent of structurally and 

909 pharmacologically comparable products for which there is clinical experience. Guidance for 

910 the pre-clinical testing of various biological products is available (Refs. 14-16). Additional 

911 consideration given to pre-clinical testing of the bioengineered pharmaceutical plant source 

912 material includes the presence and identity of potentially harmful constituents such as: 

913 toxicants, pathogens, pesticides, herbicides, fungicides, heavy metals, anti-nutrients, and 

914 allergens. Both in vitro and in vivo studies may contribute to this characterization.

915

916 For plant lines derived from a host plant or related species having a known potential to 

917 produce toxins, anti-nutrients, or allergens, you should perform sensitive tests early in 
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918 product development to demonstrate whether the levels of these components have changed 

919 in the bioengineered source plant. If the donor of the DNA is known to be a source of 

920 allergens or toxicants, then you should perform appropriate allergenicity or toxicity testing.

921

922 B. Evaluation of Impurities

923

924 Impurities and contaminants include: source-plant-derived impurities, pesticides, herbicides, 

925 fungicides, bacterial or fungal-derived impurities, and downstream processing-derived 

926 impurities. Product-related impurities include degradation products, aggregates, or other 

927 modified forms of the desired product (e.g., deamidated, isomerized, mismatched disulfide928 linked, oxidized, or altered conjugated forms). You should give special attention to post929 translational modifications unique to plant expression systems, for example the presence of 

930 xylose in glycoproteins.

931

932 Further information on this topic is provided in the ICH; Technical Requirements for 

933 Registration of Pharmaceuticals for Human Use - Guideline Q6B Specifications: Test 

934 Procedures and Acceptance Criteria for Biotechnological/Biological Products (Ref. 9).

935

936 1. Toxicants

937

938 If the host species is known to contain toxicants (e.g., protease inhibitors, 

939 hemolytic agents, neurotoxins), analytical tests, animal tests, or validation of 

940 removal may be appropriate to establish that the toxicant levels are in a safe range 

941 in the final product. Consult with FDA for further guidance.

942

943 2. Evaluation of Pesticide, Herbicide, and Fungicide Levels

944

945 You should use only pesticides, herbicides and/or fungicides registered by the 

946 Environmental Protection Agency (EPA) for use on the crop you are using. With 

947 regard to the final pharmaceutical product, you should specify the maximum 

948 amount of any pesticide, herbicide, and/or fungicide residues anticipated to be 

949 present, justify the safety of those amounts under conditions of anticipated use of 

950 the pharmaceutical, and demonstrate that the final product does not exceed those 

951 limits. A developer who has a new plant that expresses both a bioengineered 

952 biologic product and a bioengineered pesticide should consult with EPA regarding 

953 the safety of the pesticide. In some instances, validation of removal of the 

954 pesticide from the preparation may be an acceptable alternative to final product 

955 safety tests. This document only addresses FDA and USDA guidance; if you 

956 have questions regarding the use or safety of pesticides, you should contact EPA.

957

958 3. Evaluation of Metal Toxicants

959

960 You should evaluate both the presence and levels of toxic heavy metals. 

961 Consideration should be given to the host plant and whether it stores or 

962 accumulates these metals.

963
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964 C. Allergenicity 

965

966 As part of the pre-clinical evaluation, you should consider the allergenicity or 

967 immunogenicity of the intended biological product or drug. Appropriate testing protocols 

968 depend upon the intended effect of the product, the intended use (route of administration of 

969 the product), and the purity of the product. You should assess the need for allergenicity 

970 testing for each product on an individual basis and take into account production methods 

971 that might introduce allergens into the final product (e.g., from inadvertent contamination by 

972 mold, animal dander, animal excrement, or dust mite due to field or storage conditions), in 

973 addition to the potential allergenicity of the bioengineered pharmaceutical plant, itself. 

974 Consult with FDA for further guidance.

975

976 If the source plant producing the product is allergenic or immunogenic, you should test the 

977 product for those substances. Consideration should be given to plant-specific modifications, 

978 such as altered glycosylation (e.g., xylose), with regard to potential effects on immunogenic 

979 and allergenic responses to the intended product.

980

981 You should evaluate the final product for allergenic determinants, such as N-glycans. 

982 Specific serum screening of the expressed protein could be evaluated using sera derived 

983 from patients allergic to the source material. Any positive outcome from specific serum 

984 screening would define the product as likely to be allergenic.

985

986 D. Immunogenicity 

987

988 You should evaluate your product for plant specific modifications that may contribute to 

989 unintended immunogenicity. Standard immunogenicity testing for these products should be 

990 performed according to existing guidance (Refs. 14, 15) and consultation with FDA. 

991

992

993 VI. CLINICAL TESTING FOR FDA-REGULATED PRODUCTS AND PRE994 LICENSE TESTING FOR USDA-REGULATED PRODUCTS

995

996 We recommend that you refer to existing guidance(s) for conduct of clinical studies for drugs 

997 and biologics for humans and contact CDER or CBER, respectively if you have further 

998 questions. The potential residues of animal drugs (derived from bioengineered plants) in edible 

999 food animal tissues may be of concern, and you should contact CVM directly for guidance. You 

1000 should contact CVM or CVB before animal drugs or veterinary biologics are tested on non1001 laboratory animals. 

1002

1003
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1003 VII. DEFINITIONS

1004

1005 APHIS – Animal and Plant Health Inspection Service of the USDA.

1006

1007 Batch – a specific quantity of a drug or other material that is intended to have uniform character 

1008 and quality, within specified limits, and is produced according to a single manufacturing order

1009 during the same cycle of manufacture.

1010

1011 Bioengineered pharmaceutical plant – a plant manipulated by recombinant DNA technology 

1012 to express a gene encoding a biologic or drug product.

1013

1014 BLA – Biologics License Application.

1015

1016 BRS – Biotechnology Regulatory Services Division of the USDA/APHIS.

1017

1018 CFR – Code of Federal Regulations.

1019

1020 Coding region – protein coding regions contain an open reading frame which can be transcribed 

1021 into messenger RNA to direct the synthesis of a protein product.

1022

1023 Confinement – measures implemented to control the co-mingling of bioengineered 

1024 pharmaceutical plants with non-bioengineered plants or to limit the distribution of an introduced 

1025 gene to a defined area.

1026

1027 Construct – an engineered DNA fragment that contains, but is not limited to, the DNA 

1028 sequences to be integrated into a target plant's genome.

1029

1030 CBER – Center for Biologics Evaluation and Research of the FDA.

1031

1032 CDER – Center for Drug Evaluation and Research of the FDA.

1033

1034 CDRH – Center for Devices and Radiological Health of the FDA.

1035

1036 CFSAN – Center for Food Safety and Applied Nutrition of the FDA

1037

1038 CVB – Center for Veterinary Biologics of the USDA/APHIS.

1039

1040 CVM – Center for Veterinary Medicine of the FDA.

1041

1042 Direct delivery systems – gene delivery systems that do not use biological agents to introduce 

1043 foreign genes into plants. Examples include electroporation, the chemical polyethylene glycol, 

1044 microprojectile bombardment, and injection via a capillary tube or pipette.

1045

1046 Drug – human protein drug and new animal drug.

1047

1048 FDA – United States Food and Drug Administration.
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22

1049

1050 Genetic stability – the ability of the introduced DNA to be inherited in a predictable fashion and 

1051 the introduced trait to be expressed in the transformed plant line and plant lines derived 

1052 therefrom in a consistent, reliable, and predictable manner.

1053

1054 Host Plant – the parent plant prior to insertion of the gene encoding the regulated product.

1055

1056 ICH – International Conference on Harmonisation.

1057

1058 IDE – Investigational Device Exemption.

1059

1060 INAD – notice of claimed investigational exemption for a New Animal Drug that must be 

1061 submitted prior to shipment of a new animal drug for clinical tests; establishes an Investigational 

1062 New Animal Drug file, if one has not already been established for the new animal drug.

1063

1064 IND – Investigational New Drug Application.

1065

1066 Indirect delivery systems – indirect delivery systems use a biologic agent to introduce the 

1067 foreign genes into the plant's genome.

1068

1069 Lot – a batch, or a specific identified portion of a batch, having uniform character and quality 

1070 within specified limits; or, in the case of a process, it is a specific identified amount produced in 

1071 a unit of time or quantity in a manner that assures its having a uniform character and quality 

1072 within specified limits.

1073

1074 Marketing application – a BLA, NDA, NADA, PMA, 510(k), or VBPLA.

1075

1076 MSB – Master Seed Bank (or Master Seed for veterinary biologics).

1077

1078 NADA – New Animal Drug Application.

1079

1080 NDA – New Drug Application.

1081

1082 NEPA – National Environmental Policy Act.

1083

1084 New animal drug – are articles other than food intended for therapeutic, preventative, 

1085 mitigation or diagnostic purposes OR alter the structure and function of the animal.

1086

1087 Non-coding region– DNA sequences that lie outside of an open reading frame and which are 

1088 not translated to become part of a protein. These might include scaffold attachment regions, 

1089 promoters, leader sequences, enhancers, introns, terminators, and any other sequences that are 

1090 used for gene expression either in the plant or other hosts.

1091

1092 Outline of Production – a detailed protocol of methods of manufacture to be followed in the 

1093 preparation of a veterinary biological product.

1094
Draft – Not for Implementation

23

1095 Raw agricultural commodity – any food in its raw or natural state, including all unprocessed 

1096 fruits, vegetables, nuts, and grains.

1097

1098 Regulated products – FDA- or CVB-regulated intermediates, and biological products, vaccines, 

1099 and drugs, intended for human or animal use and/or animal feed.

1100

1101 Serials – consecutive lots or batches in support of a CVB product license application.

1102

1103 Source material – plant biomass from which the regulated product is produced.

1104

1105 Source plant – bioengineered host plant.

1106

1107 Source plant material – any biomass, including seeds, from a source plant.

1108

1109 Target gene – the gene encoding the regulated product, including any linked regulatory elements 

1110 and selectable markers.

1111

1112 Trait(s) – the phenotypic characteristic(s) conferred to the recipient plant by the introduced 

1113 DNA.

1114

1115 Transfection system – a method for transitory gene expression using a plant virus.

1116

1117 Transformation event – the introduction into an organism of genetic material that has been 

1118 manipulated in vitro. For the purpose of this document, ‘organism’ refers to plants.

1119

1120 Transformation system– a method for introducing new genes into plants by either direct or 

1121 indirect delivery systems.

1122

1123 USDA – United States Department of Agriculture.

1124

1125 VBPLA – United States Veterinary Biological Product License Application.

1126

1127 Vector – an autonomously replicating DNA molecule into which foreign DNA is inserted and 

1128 then propagated in a host cell.

1129

1130 Veterinary biologic - all viruses, serums, toxins, or analogous products at any stage of 

1131 production, shipment, distribution, or sale, which are intended for the use in the treatment of 

1132 animals and which act primarily trough the direct stimulation, supplementation, enhancement, or 

1133 modulation of the immune system or immune response.

1134

1135 Viral vector – a virus that has been modified to contain foreign genes.

1136

1137 Virus – infectious agents containing only nucleic acid and a protein coat that can enter and 

1138 replicate in a cell.

1139

1140 WSB – Working Seed Bank (or Working Seed for veterinary biologics).

1141
Draft – Not for Implementation

24

1141 VIII. REFERENCES

1142

1143 1. ICH; Technical Requirements for Registration of Pharmaceuticals for Human Use - Guideline 

1144 Q5B: Quality of Biotechnological Products: Analysis of the Expression Construct in Cells Used 

1145 for Production of r-DNA Derived Protein Products – (1996).

1146

1147 2. FDA Guidance for Industry: Environmental Assessment of Human Drug and Biologics 

1148 Applications – (1998).

1149

1150 3. FDA Guidance for Industry: Environmental Impact Assessments (EIA's) for Veterinary 

1151 Medicinal Products (VMP's) - Phase I – (2001).

1152

1153 4. FDA Draft Guidance for Industry: Cooperative Manufacturing Arrangements for Licensed 

1154 Biologics – (1999).

1155

1156 5. The Pesticide Product Information System contains information concerning all pesticide 

1157 products registered in the United States. (http://www.epa.gov/opppmsd1/PPISdata/index.html).

1158

1159 6. Guide to minimize microbial food safety hazards for fresh fruits and vegetables, 

1160 http://www.cfsan.fda.gov/~dms/guidance.html or 

1161 http://www.foodsafety.gov/~dms/prodguid.html

1162

1163 7. FDA Guideline on Sterile Drug Products Produced by Aseptic Processing – (1987).

1164

1165 8. FDA Guidance for Industry: For the Submission of Documentation for Sterilization Process 

1166 Validation in Applications for Human and Veterinary Drug Products – (1994).

1167

1168 9. ICH; Technical Requirements for Registration of Pharmaceuticals for Human Use - Guideline 

1169 Q6B Specifications: Test Procedures and Acceptance Criteria for Biotechnological/Biological 

1170 Products – (1999).

1171

1172 10. FDA Points to Consider in the Manufacture and Testing of Monoclonal Antibody Products 

1173 for Human Use – (1997). 

1174

1175 11. FDA Guidance for Industry: Stability Guidelines. CVM Guidance #5. (1990).

1176

1177 12. FDA Draft Guidance for Industry: Stability Testing of Drug Substances and Drug Products –

1178 (1998).

1179

1180 13. ICH; Technical Requirements for Registration of Pharmaceuticals for Human Use -

1181 Guideline Q1A(R): Stability Testing of New Drug Substances and Products – (2001).

1182

1183 14. ICH; Technical Requirements for Registration of Pharmaceuticals for Human Use -

1184 Guideline S5B: Detection of Toxicity to Reproduction for Medicinal Products – (2000).

1185
Draft – Not for Implementation

25

1186 15. ICH; Technical Requirements for Registration of Pharmaceuticals for Human Use -

1187 Guideline S6: Pre-Clinical Testing of Biotechnology-Derived Pharmaceuticals – (1997).

1188

1189 16. ICH; Technical Requirements for Registration of Pharmaceuticals for Human Use -

1190 Guideline M3: Nonclinical Safety Studies for the Conduct of Human Clinical Trials for 

1191 Pharmaceuticals – (2000).

1192

1193
Draft – Not for Implementation

26

1193 APPENDIX A

1194

1195 CONTACTS:

1196

1197 To apply for a permit for importation, interstate movement, and field testing of bioengineered 

1198 plants and plant viruses: 

1199 James White, Ph.D.

1200 U.S. Department of Agriculture

1201 Animal and Plant Health Inspection Service

1202 Biotechnology Regulatory Services, Unit 147

1203 4700 River Road 

1204 Riverdale, MD 20737

1205 Ph. # (301) 734-5940

1206 http://www.aphis.usda.gov/ppq/biotech

1207

1208 For permission to ship experimental veterinary biological products (9 CFR 103.3 authorization)

1209 or for information regarding veterinary biologics:

1210 U.S. Department of Agriculture

1211 Animal and Plant Health Inspection Service

1212 Center for Veterinary Biologics 

1213 Licensing and Policy Development 

1214 510 S. 17th St., Suite 104 

1215 Ames, Iowa 50010

1216 Ph. # (515) 232-5785; Fax # (515) 232-7120 

1217 http://www.aphis.usda.gov/vs/cvb/

1218

1219 For permission to import veterinary biological products: 

1220 U.S. Department of Agriculture

1221 Animal and Plant Health Inspection Service

1222 Center for Veterinary Biologics 

1223 4700 River Road, Unit 148 

1224 Riverdale, MD 20737

1225 Ph. # (301) 734-8245; Fax # (301) 734-4314

1226 http://www.aphis.usda.gov/vs/cvb/

1227

1228 For information regarding therapeutic or diagnostic biologics for use in humans:

1229 U.S. Food and Drug Administration

1230 Center for Biologics Evaluation and Research

1231 Office of Therapeutics Research and Review

1232 1401 Rockville Pike

1233 Rockville, MD 20852

1234 Ph. # (301) 827-5101; Fax # (301) 827-5397

1235 www.fda.gov/cber

1236

1237 For information regarding vaccines for use in humans:

1238 U.S. Food and Drug Administration
Draft – Not for Implementation

27

1239 Center for Biologics Evaluation and Research

1240 Office of Vaccines Research and Review

1241 1401 Rockville Pike

1242 Rockville, MD 20852

1243 Ph. # (301) 827-3070; Fax # (301) 827-3532

1244 www.fda.gov/cber

1245

1246 For information regarding animal feeds and animal drugs:

1247 U.S. Food and Drug Administration

1248 Center for Veterinary Medicine

1249 HFV-200, 7500 Standish Place

1250 Rockville, MD 20855

1251 Ph. # (301) 827-6652; Fax # (301) 827-1484 

1252 www.fda.gov/cvm

1253

1254 For consultation on issues related to human food:

1255 U.S. Food and Drug Administration

1256 Center for Food Safety and Applied Nutrition

1257 HFS-013, 5100 Paint Branch Parkway

1258 College Park, MD 20740-3835 

1259 Ph # (301) 436-1715; Fax # (301) 436-2637

1260 www.cfsan.fda.gov

1261

1262 For information regarding drugs for use in humans:

1263 U.S. Food and Drug Administration

1264 Center for Drug Evaluation and Research 

1265 5600 Fishers Lane

1266 Rockville, MD 20857

1267 Ph.# (301) 827-4573; Fax # (301) 827-3056

1268 www.fda.gov/cder

1269

1270 For information regarding medical devices:

1271 U.S. Food and Drug Administration

1272 Center for Devices and Radiological Health 

1273 Division of Small Manufacturers Assistance

1274 1350 Piccard Drive

1275 Rockville, MD 20850

1276 Ph.# (301) 443-6597; Fax # (800) 638-2041

1277 www.fda.gov/cdrh