Federal Register / Vol. 60, No. 95 / Wednesday, May 17, 1995 / Proposed Rules 26381

prior to moving interstate by attaching

to the left ear a metal tag bearing a serial

number and the inscription ‘‘U.S.

Reactor,’’ or a metal tag bearing a serial

number designated by the State animal

health official for identifying brucellosis

reactors, and must be:

(1) ‘‘B’’ branded (as defined in § 78.1);

or

(2) Accompanied directly to slaughter

by an APHIS or State representative; or

(3) Moved in vehicles closed with

official seals applied and removed by an

APHIS representative, State

representative, accredited veterinarian,

or an individual authorized for this

purpose by an APHIS representative.

The official seal numbers must be

recorded on the accompanying permit.

* * * * *

Done in Washington, DC, this 11th day of

May 1995.

Lonnie J. King,

Acting Administrator, Animal and Plant

Health Inspection Service.

[FR Doc. 95–12150 Filed 5–16–95; 8:45 am]

BILLING CODE 3410–34–P

9 CFR Parts 101 and 113

[Docket No. 94–051–1]

RIN 0579–AA66

Viruses, Serums, Toxins, and

Analogous Products; In Vitro Tests for

Serial Release

AGENCY: Animal and Plant Health

Inspection Service, USDA.

ACTION: Proposed rule.

SUMMARY: We are proposing to amend

the regulations regarding the use of in

vitro potency tests in place of animal

tests for immunogenicity to: change the

title of the section; prescribe

requirements for in vitro immunoassays

used to determine the relative antigen

content of inactivated biological

products; require that such

immunoassays be parallel line assays

based upon unexpired reference

preparations; require that in vitro tests

for relative antigen content be converted

to parallel line assays within 2 years;

specify procedures and requirements for

qualifying or requalifying reference

preparations for inactivated products;

and add certain definitions to the

regulations.

The effect of the amendment would

be to standardize the methods used to

determine the relative potency of

inactivated biological products.

DATES: Consideration will be given only

to comments received on or before

August 15, 1995.

ADDRESSES: Please send an original and

three copies of your comments to

Docket No. 94–051–1, Regulatory

Analysis and Development, PPD,

APHIS, Suite 3C03, 4700 River Road

Unit 118, Riverdale, MD 20737–1238.

Please state that your comments refer to

Docket No. 94–051–1. Comments

received may be inspected at USDA,

room 1141, South Building, 14th Street

and Independence Avenue SW.,

Washington, DC, between 8 a.m. and

4:30 p.m., Monday through Friday,

except holidays. Persons wishing to

inspect comments are requested to call

ahead (202) 690–2817 to facilitate entry

into the comment reading room.

FOR FURTHER INFORMATION CONTACT:

Dr. David A. Espeseth, Deputy Director,

Veterinary Biologics, BBEP, APHIS,

4700 River Road Unit 148, Riverdale,

MD 20737–1237, (301) 734–8245.

SUPPLEMENTARY INFORMATION:

Background

The regulations pertaining to the

testing of biologics provide that no

biological product shall be released (for

sale) prior to the completion of tests

prescribed to establish the product to be

pure, safe, potent, and efficacious (9

CFR 113.5). Efficacy refers to the

specific ability of the product to effect

the result for which it is offered when

used as recommended by the

manufacturer. Studies conducted to

establish efficacy include

immunogenicity tests in host animals

using product manufactured according

to specific requirements which include

specifications for antigen content and/or

animal potency. These requirements

apply to every serial of product which

is produced. Therefore, if a product has

been tested for immunogenicity in

animals and shown to have the desired

effect, it follows that subsequent serials

(batches) of the product manufactured

to the same specifications should also

have the same effect.

Once immunogenicity is established

in relation to a specific minimum

antigen content in a product, it should

no longer be necessary to test every

subsequent serial for potency in animals

if an evaluation can be made with

reasonable certainty of the relative

antigen content by testing the serial or

subserial in an acceptable in vitro test

system. Therefore, when properly

qualified and validated, in vitro

immunoassays that determine relative

antigen content of the product can serve

as acceptable substitutes for potency

tests that otherwise would need to be

performed in animals.

The regulations in 9 CFR 113.8

pertain to the use of in vitro tests in

place of animal tests for determining the

potency of veterinary biological

products. Currently, the in vitro tests

prescribed in § 113.8 which include

determining the log10 virus titer and

performing live bacterial counts are only

applicable to veterinary biologicals

which contain live microorganisms. The

changes and test procedures prescribed

in this proposal would make § 113.8

applicable to both live and inactivated

products by prescribing validity

requirements for in vitro test systems

used, in place of animal tests, to test for

the potency of inactivated products.

We are proposing to amend the title

of § 113.8 to read: ‘‘In vitro potency tests

for serial release.’’ This change is

intended to clarify the fact that the in

vitro procedures described in § 113.8 are

applicable to in vitro tests used to

release serials or subserials of veterinary

biological products after the prescribed

animal protection studies required for

licensing have been completed. In the

case of inactivated products, the

proposal specifies that in vitro

immunoassays (test systems) which

compare the relative antigen content

(relative potency) of a test serial to a

reference preparation must be parallel

line assays using an unexpired reference

preparation whose potency has been

correlated directly or indirectly to

immunogenicity in host animals.

In addition, the proposal would

require: confirming the accuracy of the

protective dose established for live

products 3 years after the initiation of

the host animal immunogenicity study;

and confirming the immunogenicity of

reference preparations used in

immunoassays for inactivated biological

products prior to their expiration. The

expiration date for a reference would be

equal to the dating of the product or as

supported by data acceptable to APHIS.

APHIS is proposing these

amendments because current

requirements for many of the

immunoassays being used to release

serials or subserials of product do not

have uniform validity criteria and do

not include a provision to confirm

periodically the immunogenicity of the

reference used in such immunoassays.

The proposed amendment would

standardize the requirements for in vitro

potency tests for relative antigen content

and update and improve the reliability

of such tests that are currently included

in filed outlines of production. The

proposed amendment does not specify a

particular immunoassay provided that it

is a parallel line assay using an

unexpired reference preparation. While

there is not a generally accepted ‘‘best’’

immunoassay, there is general

agreement that an acceptable
26382 Federal Register / Vol. 60, No. 95 / Wednesday, May 17, 1995 / Proposed Rules

immunoassay must demonstrate

linearity, specificity, and

reproducibility; and that the reference

must be capable of eliciting a protective

immune response in animals for as long

as it serves as the reference.

APHIS has selected the parallel line

assay because it demonstrates linearity,

specificity, and reproducibility, and also

compares the ‘‘similarity’’ of the

responses elicited by the test and

reference preparations in the

immunoassay. APHIS feels that the

‘‘similarity’’ feature is critical to the

acceptance of any in vitro immunoassay

purporting to measure relative antigenic

content. We realize that the proposed

amendment would necessitate the

revalidation of immunoassays currently

contained in some filed outlines of

production and propose to implement

the requirements as set out below.

Firms with filed outlines of

production for licensed products with

in vitro potency tests that are

immunoassays that are not parallel line

assays would be allowed 2 years after

the effective date of the final rule to

come into compliance with the

proposed amendments. In the interim,

immunoassays, utilizing unexpired

references, contained in previously

approved outlines of production would

continue to be allowed for serial release

of previously licensed fractions but

would not be acceptable for fractions

not previously licensed to the firm.

Firms with filed outlines of production

for licensed product with in vitro

potency tests for relative antigen content

would be required to use unexpired

references. References that have expired

or that are about to expire would need

to be requalified or have the dating

period extended in accordance with

protocols and time schedules acceptable

to APHIS.

APHIS has determined that

immunoassays that are not based on a

parallel line assay using an unexpired

reference may not provide reliable

relative potency data in all instances.

Such instances include the

determination of relative potency based

on a reference preparation that may

have expired or the extrapolation of data

based upon standard curves that may

not be proportional between serial and

reference. These amendments are being

proposed in order to provide greater

assurance that a serial of product

provides adequate potency in all

instances.

We are also adding to the regulations

in § 101.5 definitions of the term

‘‘immunogenicity’’, and the terms

‘‘master reference’’, ‘‘working

reference’’, and ‘‘qualifying serial’’ as

they apply to reference preparations

used in in vitro immunoassays.

Licensees, researchers, and scientists

at the National Veterinary Services

Laboratories, U.S. Department of

Agriculture, have cooperated in the

development of this proposed rule. We

are therefore proposing to amend

§§ 101.5 and 113.8 as set forth below.

Executive Order 12866 and Regulatory

Flexibility Act

This proposed rule has been reviewed

under Executive Order 12866. The rule

has been determined to be no significant

for purposes of Executive Order 12866,

and therefore, has not been reviewed by

the Office of Management and Budget.

This proposed amendment, if

adopted, would allow any valid in vitro

immunoassay to be used in determining

the relative antigen content of an

inactivated veterinary biological

product, provided that it satisfies the

parallel line criteria and that it is

conducted using an unexpired reference

preparation that has been tested,

directly or indirectly, for

immunogenicity in a manner acceptable

to APHIS. This amendment would affect

all licensed manufacturers of veterinary

biologicals utilizing in vitro relative

potency immunoassays for determining

the potency of inactivated products.

This proposal, however, does not

impose any additional economic burden

since the testing of product for potency

is already required under § 113.5 of the

regulations and outlines of production

are routinely amended and updated.

Section 113.5 specifies that no

biological product shall be released

prior to the completion of tests

prescribed in a filed outline of

production or standard requirement to

establish that the product is pure, safe,

potent, and efficacious. In the absence

of a standard requirement prescribing a

specific potency test for inactivated

products, the firms develop a potency

test suitable for their product, and

designate such tests in the outline of

production that is filed with APHIS.

Currently, firms are using host animal

tests, laboratory animal tests, and a

variety of in vitro immunoassays as

potency tests for inactivated products.

This proposed rule does not restrict the

firm’s discretion to choose the most

appropriate test for its product. The

proposed rule would only prescribe

validity requirements for in vitro

immunoassays for relative potency. The

overall effect of this proposed

amendment would be to standardize in

vitro immunoassays that are used to

determine the potency of inactivated

veterinary biological products.

Under these circumstances, the

Administrator of the Animal and Plant

Health Inspection Service has

determined that this action would not

have a significant economic impact on

a substantial number of small entities.

Executive Order 12372

This program/activity is listed in the

Catalog of Federal Domestic Assistance

under No. 10.025 and is subject to

Executive Order 12372, which requires

intergovernmental consultation with

State and local officials. (See 7 CFR part

3015, subpart V.)

Executive Order 12778

This proposed rule has been reviewed

under Executive Order 12778, Civil

Justice Reform. If this proposed rule is

adopted: (1) All State and local laws and

regulations that are in conflict with this

rule will be preempted; (2) no

retroactive effect will be given to this

rule; and (3) administrative proceedings

will not be required before parties may

file suit in court challenging this rule.

Paperwork Reduction Act

In accordance with the Paperwork

Reduction Act of 1980 (44 U.S.C. 3501

et seq.), the information collection or

recordkeeping requirements included in

this proposed rule have been approved

by the Office of Management and

Budget (OMB), and there are no new

requirements. The assigned OMB

control number is 0579–0013.

List of Subjects

9 CFR Part 101

Animal biologics.

9 CFR Part 113

Animal biologics, Exports, Imports,

Reporting and recordkeeping

requirements.

Accordingly, 9 CFR parts 101 and 113

would be amended as follows:

PART 101—DEFINITIONS

1. The authority citation for part 101

would continue to read as follows:

Authority: 21 U.S.C. 151–159; 7 CFR 2.17,

2.51, and 371.2(d).

2. Section 101.5 would be amended

by adding new paragraphs (o), (p), (q),

and (r) to read as follows:

§ 101.5 Testing terminology.

* * * * *

(o) Master Reference. A Master

Reference is a reference whose potency

is correlated, directly or indirectly, to

host animal immunogenicity. The

Master Reference may be used as the

working reference in in vitro tests for

relative potency. The Master Reference
Federal Register / Vol. 60, No. 95 / Wednesday, May 17, 1995 / Proposed Rules 26383

may also be used to establish the

relative potency of a serial of product

used in requalification studies and to

establish the relative potency of working

references. A Master Reference may be:

(1) A completed serial of vaccine or

bacterin prepared in accordance with a

field Outline of Production;

(2) A purified preparation of the

protective immunogen or antigen; or

(3) A nonadjuvanted harvested

culture of microorganisms.

(p) Working Reference. A Working

Reference is the reference preparation

that is used in the in vitro test for the

release of serials of product. Working

References may be:

(1) Master References; or

(2) Serials of product that have been

prepared and qualified, in a manner

acceptable to APHIS, for use as

reference preparations.

(q) Qualifying Serial. (1) A serial of

biological product used to test for

immunogenicity when the Master or

Working Reference is a purified antigen

or nonadjuvanted harvest material.

Qualifying serials shall be produced in

accordance with the filed Outline of

Production, tested for immunogenicity

in host animals in accordance with

protocols acceptable to Animal and

Plant Health Inspection Service, and

have a geometric mean relative potency,

when compared to the Master

Reference, of not greater than 1.0 as

established by independent parallel line

assays with 5 or more replicates.

(2) Qualifying serials used to requalify

or extend the dating period of a Master

Reference in a repeat immunogenicity

test shall satisfy all criteria prescribed

above and, it addition, shall have been

prepared within 6 months of

requalifying testing, i.e., the initiation of

the repeat immunogenicity test.

(r) Immunogenicity. The ability of a

biological product to elicit an immune

response in animals as determined by

test methods or procedures acceptable

to the Animal and Plant Health

Inspection Service.

PART 113—STANDARD

REQUIREMENTS

3. The authority citation for part 113

would continue to read as follows:

Authority: 21 U.S.C. 151–159; 7 CFR 2.17,

2.51, and 371.2(d).

4. Section 113.8 would be amended as

follows:

a. The section heading would be

revised to read as set forth below.

b. Paragraph (a) would be revised to

read as set forth below.

c. Paragraph (b) the introductory text

would be revised to read as set forth

below.

d. Paragraph (b)(5) would be revised

to read as set forth below.

e. Paragraph (c) would be

redesignated as paragraph (e) and new

paragraphs (c) and (d) would be added

to read as set forth below.

f. In redesignated paragraph (e), in the

introductory text, the reference to

‘‘paragraph (b)’’ would be removed and

‘‘Paragraphs (b) and (c)’’ would be

added in its place. In paragraph (e)(4),

the reference to ‘‘paragraphs (c)(1),’’

would be removed and ‘‘paragraphs

(e)(1),’’ would be added in its place.

§ 113.8 In vitro potency tests for serial

release.

(a) Master Seed which has been

established as pure, safe, and

immunogenic shall be used for

preparing seed for production as

specified in the Standard Requirements

or in the filed Outline of Production.

The Administrator may exempt a

product from a required animal potency

test for release when an evaluation can,

with reasonable certainty, be made by:

(1) Subjecting the master seed to the

applicable requirements prescribed in

§§ 113.64, 113.100, 113.200, and

113.300;

(2) Testing the Master Seed for

immunogenicity in a manner acceptable

to the Animal and Plant Health

Inspection Service (APHIS);

(3) Establishing a satisfactory potency

for live products based on the protective

dose used in the Master Seed

immunogenicity test plus an adequate

overage allowance for adverse

conditions and test error; and

(4) For inactivated products,

determining the potency of each serial

or subserial, or both, using an accepted

test system. Acceptable potency tests

shall include:

(i) Determining the log10 live virus

titer;

(ii) Determining the live bacterial

count; or

(iii) For inactivated products,

determining the relative antigen

content, as compared with a reference,

using a parallel line immunoassay.

(b) In the case of live products, each

serial and subserial of desiccated

product derived from an approved

Master Seed and bulk or final container

samples of each serial of completed

liquid product derived from an

approved Master Seed shall be

evaluated by a test procedure acceptable

to APHIS. On the basis of the results of

the test, as compared with the required

minimum potency, each serial and

subserial shall either be released to the

firm for marketing or withheld from the

market. The evaluation of such products

shall be made in accordance with the

following criteria:

(1) * * *

* * * * *

(5) Exceptions. When a product is

evaluated in terms other than log10 virus

titer or organism count, an appropriate

difference between the average potency

value obtained in the retests and the

potency value obtained in the initial test

shall be established for use in

paragraphs (b)(3) or (b)(4) of this section

to evaluate such products and shall be

specified in the Product Standard

Requirement or filed Outline of

Production.

(c) In the case of inactivated products,

bulk or final container samples of

completed product from each serial

derived from an approved Master Seed,

shall be evaluated for relative antigen

content (potency), as compared with a

reference, by a parallel line

immunoassay procedure acceptable to

APHIS. Firms currently using

immunoassays which do not meet the

requirements of a parallel line assay

shall have 2 years from the effective

date of the final rule to update their

filed Outlines of Production to be in

compliance with this requirement. On

the basis of the results of such test

procedures, each serial that meets the

required minimum potency shall be

released to the firm for marketing; each

serial not meeting the required

minimum potency shall be withheld

from the market. The evaluation of such

products shall be made in accordance

with the following criteria:

(1) A test that results in no valid lines

is considered a no test and may be

repeated.

(2) An initial test that results in valid

lines that are not parallel is considered

a valid equivocal test. Release of the

serial may not be based on such test

since the result cannot be termed

‘‘satisfactory’’ or ‘‘unsatisfactory’’.

(3) If the initial test shows that

potency equals or exceeds the required

minimum potency, the serial is

satisfactory without additional testing.

(4) If the initial test is an equivocal

test due to lack of parallelism, the serial

may be retested up to three times:

Provided, That, if the test is not

repeated, the serial shall be deemed

unsatisfactory.

(i) If more than 50% of all valid repeat

tests show that potency equals or

exceeds the required minimum potency,

the serial is satisfactory.

(ii) If greater than 50% of all valid

repeat tests show either lack of

parallelism or that potency is less than

the required minimum potency, the

serial is unsatisfactory.
26384 Federal Register / Vol. 60, No. 95 / Wednesday, May 17, 1995 / Proposed Rules

(5) If the initial test shows that

potency is less than the required

minimum potency, the serial may be

retested. If retested, two additional tests,

must be conducted: Provided, That, if

the serial is not retested, the serial shall

be deemed unsatisfactory.

(i) If more than 50% of all valid tests

show that potency equals or exceeds the

required minimum potency, the serial is

satisfactory.

(ii) If more than 50% of all valid tests

show either lack of parallelism or that

potency is less than the required

minimum potency, the serial is

unsatisfactory.

(d) Repeat immunogenicity tests.

(1) The accuracy of the protective

dose established for live products in the

Master Seed immunogenicty test and

defined as live virus titer or live

bacterial count shall be confirmed in 3

years in a manner acceptable to APHIS,

unless use of the lot of Master Seed

previously tested is discontinued.

(2) All determinations of relative

antigen content using parallel line

immunoassays shall be conducted with

an unexpired reference. The lot of

reference used to determine antigenic

content shall have an initial dating

period equal to the dating of the product

or as supported by data acceptable to

APHIS. Prior to the expiration date,

such reference may be granted an

extension of dating by confirming its

immunogenicity using a Qualifying

Serial of product. Tests to establish or

confirm immunogenicity of references

shall be conducted in a manner

acceptable to APHIS. The dating period

of the Master Reference and Working

Reference may be extended as

supported by data acceptable to APHIS

if the minimum potency of the Master

Reference is determined to be

adequately above the minimum level

needed to provide protection in the host

animal. If a new Master Reference is

established, it shall be allowed an initial

dating period equal to the dating of the

product or as supported by data

acceptable to APHIS.

* * * * *

Done in Washington, DC, this 11th day of

May 1995.

Terry L. Medley,

Acting Administrator, Animal and Plant

Health Inspection Services.

[FR Doc. 95–12152 Filed 5–16–95; 8:45 am]

BILLING CODE 3410–34–M

9 CFR Part 113

[Docket No. 93–039–2]

Viruses, Serums, Toxins, and

Analogous Products; Standard

Requirement or Escherichia Coli

Bacterin

AGENCY: Animal and Plant Health

Inspection Service, USDA.

ACTION: Notice of reopening and

extension of comment period.

SUMMARY: We are reopening and

extending the comment period for the

proposed Standard Requirement for

Escherichia coli bacterin. This extension

will provide interested persons with

additional time in which to prepare

comments on the proposed rule.

DATES: Consideration will be given only

to written comments on Docket No. 93–

039–1 that are received on or before

August 15, 1995.

ADDRESSES: Please send an original and

three copies of your comments to

Docket No. 93–039–1, Regulatory

Analysis and Development, PPD,

APHIS, Suite 3C03, 4700 River Road

Unit 118, Riverdale, MD 20737–1238.

Please state that your comments refer to

Docket No. 93–039–1. Comments

received may be inspected at USDA,

room 1141, South Building, 14th Street

and Independence Avenue SW.,

Washington, DC, between 8 a.m. and

4:30 p.m., Monday through Friday,

except holidays. Persons wishing to

inspect comments are requested to call

ahead on (202) 690–2817 to facilitate

entry into the comment reading room.

FOR FURTHER INFORMATION CONTACT: Dr.

David Espeseth, Deputy Director,

Veterinary Biologics, BBEP, APHIS,

4700 River Road Unit 148, Riverdale,

MD 20737–1237, (301) 734–8245.

SUPPLEMENTARY INFORMATION: On

October 11, 1994, we published in the

Federal Register (59 FR 51390–51392,

Docket No. 93–039–1) a proposed rule

to amend the regulations in 9 CFR

113.124 to include a Standard

Requirement for Escherichia coli

bacterins. Comments on the proposed

rule were required to be received on or

before December 12, 1994.

So that we may consider comments

submitted after that date, we are

reopening and extending the public

comment period on Docket No. 93–039–

1 until 90 days after the date of

publication of this notice in the Federal

Register. During this period, interested

persons may submit their comments for

our consideration.

Authority: 21 U.S.C. 151–159, 7 CFR 2.17,

2.51, and 371.2(d). Done in Washington, DC,

this 11th day of May 1995.

Lonnie J. King,

Acting Administrator, Animal and Plant

Health Inspection Service.

[FR Doc. 95–12151 Filed 5–16–95; 8:45 am]

BILLING CODE 3410–34–M

DEPARTMENT OF TRANSPORTATION

Federal Aviation Administration

14 CFR Part 71

[Airspace Docket No. 95–ANE–23]

Proposed Establishment of Class E

Airspace; Portland, ME

AGENCY: Federal Aviation

Administration (FAA), DOT.

ACTION: Notice of proposed rulemaking.

SUMMARY: This proposed rule would

establish Class E airspace at Portland

International Jetport, Portland, Maine,

that coincides with the hours that the

associated radar approach control

facility is not in operation. Since the

Portland Class C airspace is predicated

on an operational air traffic control

tower (ATCT) serviced by a radar

control approach facility (TRACON),

Class E airspace must be defined for the

hours when that facility is not in

operation. This proposal would not

change the designated boundaries or

altitudes of the Portland Class C

airspace, but only establish the

necessary Class E airspace to provide

sufficient controlled airspace for those

aircraft operating under instrument

flight rules during the hours when the

Portland ATCT and TRACON are not in

operation.

DATES: Comments must be received on

or before June 16, 1995.

ADDRESSES: Send comments on the

proposal in triplicate to: Manager,

System Management Branch, ANE–530,

Federal Aviation Administration, 12

New England Executive Park,

Burlington, MA 01803–5299; telephone

(617) 238–7530; fax (617) 238–7596.

The official docket may be examined

in the Office of the Assistant Chief

Counsel for the New England Region,

ANE–7, 12 New England Executive

Park, Burlington, MA 01803–5299;

telephone (617) 238–7049; fax (617)

238–7055.

An informal docket may also be

examined during normal business hours

in the Office of the Manager, System

Management Branch, Air Traffic

Division, ANE–530, at the first address

shown above.