Monoclonal Antibody
Discovery Service
using Rabbit DNA Immunizationand Single B-Cell Cloning

About Us

Company Business Plan

FUJIFILM Wako Pure Chemical is a
general life science company from Japan
with almost 100 years of history. Basedon advanced technological development,
we provide highly functional andhigh-quality laboratory chemicals,
specialty chemicals, and clinicaldiagnostic reagents as our products, inaddition to cutting-edge CRO services.
With our presence in Japan and our local
subsidiaries in Europe, the US, and China,
we strive to contribute to the global life
science/chemical industry.

Business Segments
Laboratory Chemicals
Portfolio of 4000+ products for cell culture, life
sciences, and chemistry, as well as CRO services
Clinical Diagnostic Reagents
High-quality diagnostic reagents, tests, and assay kits for biochemistry and disease detection
Specialty Chemicals
Reagents for polymer chemistry, pharmaceutical/cosmetic applications, and electronics

Our CRO Services
Based on solid data and innovative researchExperienced scientists and premium customer service
Highly customizable service based on customers’ needs

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Monoclonal Antibody Discovery Company Business Plan
Using Rabbit DNA Immunization and Single B-cell Cloning

Our experienced antibody experts offer our clients a tailored
milestone-based service for antibody discovery.
Overview

By using the DNA immunization technique with rabbit monoclonal antibody production, weare able to obtain antibodies that possess both the benefits of the DNA immunization
method and the “high efficacy and affinity” properties of monoclonal rabbit antibodies.
These antibodies can be used as a next-generation antibodies for antibody therapeutics (i.e.,
TCR/CAR-T discovery, ADCs), diagnostic reagents, and more.

What is DNA immunization?

In the conventional production of
monoclonal antibodies, recombinant
proteins are derived from DNA or
peptide synthesis and thenadministered as antigens to immunized animals to generate antibodies.
In contrast, the DNA immunization
method involves directly injecting aplasmid into the animal to express
the target protein within its body. Thismethod avoids the need for antigen
proteins/peptides, prevents
contamination and denaturation, and allows easy modification of the target antigen.
Additionally, it facilitates the production of neutralizing (functional) antibodies.

Advantages of Rabbit Monoclonal Antibody•

Highly specific high affinity antibodies1••

More similar to human genome than to rodents1
Broader recognition of immunogenic antigens and epitopes2

Benefits at a Glance
Antibodies that recognize the native form of antigen of interest andchallenging membrane proteinsNo need for recombinant protein/peptide synthesis for immunization
Guaranteed high affinity and performance next-gen antibodies

1: Mage, R. G., Esteves, P. J., & Rader, C. (2019). Rabbit models of human diseases for diagnostics and therapeutics development. Developmental and Comparative Immunology, 92, 99–104. https://doi.org/10.1016/j.dci.2018.10.003
2: Weber, J., Peng, H., & Rader, C. (2017). From rabbit antibody repertoires to rabbit monoclonal antibodies. Experimental & Molecular Medicine, 49(3), e305–e305. https://doi.org/10.1038/emm.2017.23

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Milestones
Target Information(Accession No. & Application)

Feasibility Study
2 weeks

M1: Antigen Expression Analysis
1.5 months

M2: Rabbit Immunization
2.5 months

M3: mAb Production
Harvesting Splenocytes
Single B-Cell Cloning
Expression Check &Sequencing
2-3 months

Deliverables
Antibody Gene Sequence
ReportService Information

Feasibility Study
Based on the requested sequence information and intended use,
we analyze the structure and properties of the target in silico andpropose an antibody production strategy. We will propose theoptimal immunizing region, immunization method, and screening
method together with a report.
Milestone 1: Antigen Expression Analysis
Based on the Feasibility Study, target genes are cloned into vectors provided by the customer or into our inhouse expression vectors to create immunizing plasmids. After transient expression in
mammalian cells, we check antigen expression on the cell surface
using a flow cytometer.
**If the customer already has a vector that is known to express
the antigen, Milestone 1 can be omitted.
Milestone 2: Rabbit Immunization
Japanese white rabbits are injected subcutaneously with theimmunizing plasmid. Antiserum is collected after about 2 monthsof immunization. Antibody titers are evaluated by flow cytometry
using transient forced expression cells produced in Milestone 1.
Milestone 3: mAb Production
Harvesting: Spleens from rabbits with elevated antibody titers
are harvested and splenocytes are obtained. Splenocytes areseeded into plates and cells are separated one by one.
Single B-Cell Screening:
Primary screening is conducted in chambers seeded with spleencells. By coating the chamber with extracellular vesicles (EVs) thatexpress antigens on their membranes, membrane proteins canbe screened according to our Fujifilm proprietary method (PAT:WO2024071374)3. The chambers are coated with antigen, andcells that secrete antibodies binding to the antigen are picked (upto 44 cells). The antibody genes are then amplified via single-cell
PCR. In secondary screening, the amplified gene fragments aretransiently expressed in mammalian cells, and their antigen-binding capabilities are assessed using flow cytometry or ELISA.
Expression Check & Sequencing:
Six clones are selected and an expression vector carrying theantibody gene is constructed. The expression vector is transiently
expressed in mammalian cells, and antigen binding and expression levels are analyzed by flow cytometry or ELISA. We will thensequence the antibody genes and deliver the antibody genesequences and expression vectors.

3: World Intellectual Property Organization. (2024). Screening method for cell that produces objective substance, nucleic acid production method, and objective substance production method.
(WO/2024/071374). Retrieved from https://patentscope.wipo.int/search/en/detail.jsf?docId=WO2024071374

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Distribution & Inquiries
FUJIFILM Wako Chemicals Europe GmbH
Fuggerstr. 12 • 41468 Neuss, Germany
Telephone: +49-2131-311-271
Fax:+49-2131-311-110
E-mail: labchem_wkeu@fujiﬁlm.com
FUJIFILM Wako Pure Chemical Corporation
1-2, Doshomachi 3-Chome Chuo-Ku, Osaka 540-8605, Japan
Telephone: +81-6-6203-3741
Fax:+81-6-6203-1999
E-mail: ﬀwk-cservice@fujiﬁlm.com

https://labchem-wako.fujiﬁlm.com
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