• Rabbit Monoclonal Antibodies (mAbs)
    • Affordable, Comprehensive Custom Service
    • Single B Cell Express Cloning
    • Alpaca Antibody Service Available
    Rabbit Monoclonal Antibodies (mAbs)

Rabbit Monoclonal Antibody Development

Applied StemCell (ASC) is a fast-growing gene and cell therapy CRO that continuously works to expand its technologies, services, and products to address the current and future needs of the industry and research community. ASC understands that monoclonal antibodies (mAbs) are essential for research and development, so we established and optimized a single B cell antibody discovery platform that can be used to generate high-quality rabbit or alpaca antibodies. Choose from our selection of unique rabbit strains/lines, or ask us about our customized gene knockout rabbit. You can select our classical or accelerated immunization protocol based on your timeline constraints. Once the immunization process is complete, our team conducts single B cell cloning and delivers 40 ELISA positive recombinant mAbs and a data report. You can screen and select your clones of interest, or ASC can complete the screening process for you (*additional fees apply). After antibody selection, our scientists sequence and produce the elected antibodies.

During your free consultation, our experts can help you design a flexible project outline that captures all of your requirements. Once the project design is finalized, you send us your antigen and we take care of the rest. ASC will have your rabbit monoclonal antibodies in just a few weeks.

  • Rabbit Monoclonal Antibodies with Subnanomolar Affinity
  • HyImmune Rabbits typically produce 3× antigen specific B cells, to increase the antibody development rate
  • Unlimited Supply: Customer owns IP
  • More Antibodies: >0.5mg for each antibody selected
  • Convenient: Antibodies can be delivered with or without luciferase
  • Alpaca Antibody Service Available - Inquire
  • Affordable Pricing
  • Fast turnaround time

Advantages of ASC's Single B Cell Antibody Discovery Platform

Monoclonal Antibodies - Unique Animal Strains and Lines

Unique Animal Strains & Lines

- LucAb rabbit: produces antibodies with luciferase
- Hu rabbit: high antibody humanization
- HyImmune rabbit: enhanced immune response
- Customized Rabit Line: gene knockout
- Alpaca

Unique Animal Strains & Lines

Monoclonal Antibodies - Optimized Immunization Protocol

Optimized Immunization Protocol

- Accelerated protocol: 4 weeks
- Classical Protocol: 2-months
- We ensure antibody affinity & diversity
- Serum titer monitering 

Optimized Immunization Protocol

Monoclonal Antibodies - Single B Cell mRNA Amplification

Rapid Amplification of Single B Cell mRNA

- Single B Cell Sorting: 1,000 antigen-specific single B cells/day
- mRNA Extraction & Amplification: 2 days
- We guarantee monoclonality
- Transfection Ready

Rapid Amplification of Single B Cell mRNA

Monoclonal Antibodies

Efficient Mammalian Cell Antibody Expression

- First Batch: 96 antibodies in 1 week
- >1,000 antibodies/month
- ELISA positive antibodies are provided for screening
- Provided Data: Antibody concentration & ELISA binding

Efficient Mammalian Cell Antibody Expression

Rabbit Antibody Production Better than Hybridoma and Phase Display

Why Rabbit & Alpaca?

Rabbit mAbs Alpaca mAbs
  • Classic Antibody Animal
  • High Affinity: 10~100x higher than mouse mAbs
  • Strong Immune Response 
  • Cross-Reactivity
  • Custom Strain/Line Development 
  • Size: small; nanobodies (<15kd) can be made 
  • Flexible: bi-, tri- specificity
  • Express in E. coli
  • More Applications

 

Service Overview

Rabbit Monoclonal Antibody Discovery Platform

Stage 1:

Customer Provides Antigen

Stage 2:

Animal Immunization

Stage 3:

Single B Cell Cloning

Stage 4:

Customer In House Screening

Stage 5:

Antibody Sequencing & Production

  • Protein to protein conjugate
  • 3.5 mg, >0.5 mg/mL in PBS
  • 3 mg for immunization; 0.5 mg for screening
  • Accelerated Immunization (HyImmune rabbit): 1 animal, 4 weeks
  • Classical immunization: 1 animal, 9 weeks
  • Deliverable: Serum titer report
  • 2 weeks
  • Deliverables:
    • 40 ELISA positive recombinant mAbs
    • Report: mAb concentration & antigen binding
  • Select clones of interest

Additional Service: Functional Assay Development

  • 3 weeks
  • Deliverables:
    • Sequencing data: 3 selected antibodies
    • >0.5 mg of the selected 3 antibodies

Additional Services

Stage 0: 

Antigen Preparation

Stage 6:

Cell Line Generation & Assay Development

  • Peptide synthesis; 2-3 weeks
  • ≤25 Aa, 9mg, >85% purity
  • Peptide conjugation: 2 days
  • Antibody CHO cell line generation 
  • Antibody pair screening 
  • Antibody humanization

 

Deliverables 

Stage 2 Stage 3 Stage 5
  • Serum titer report
  • 40 ELISA positive recombinant mAbs
  • Report: mAb concentration & antigen binding
  • Sequencing data: 3 selected antibodies
  • >0.5 mg of the selected 3 antibodies
FAQs
Which cell surface makers did you use to sort out rabbit B cells?
Could we used SBEAD approach to generate mAb against peptide having 8 to 12 amino acids long?
Would it be possible to use your SBEAD technology to generate the 40 positive ELISA clones and continue with antibody production in our lab or do you require production of the antibody in your facility only?
What is the cost of the Ab discovery via HyImmune rabbit and SBEAD protocol?
Is the Hyimmune rabbit lacking the LC C-C bond wildtype or engineered rabbit?
What is the timeline from immunization to clone sequence delivery?
How many Ab clones are delivered for each immunization.
Does ASC do antibody humanization?
Could the client obtain the leftover B cells from the SBEAD screening and make a phage display library from it?
Can you also generate polyclonal antibodies in parallel with monoclonal antibody development?
Can SBEAD be applied to human samples?
Can antibodies be developed with low amount of antigen, e.g. <0.5mg?
Products and Services
Case Studies

Case Study 1: Identification of Antibodies for COVID-19 Antigen Detection

Rabbit Line: LucAb Rabbit

Reasons For Using the Rabbit Line: Luciferase allows quantification of antibodies and enables sandwich ELISA setup; Sandwich ELISA = used to detect ultrasensitive antibody pairs

Results: The antibody pair that we identified can be incorporated into a sandwich ELISA, and it can detect 0.5 pg/ml COVID-19 Nucleocapsid in human serum samples.

*Sensitivity is much higher than other products

Monoclonal Antibodies - COVID-19 Case Study

Figure 1: COVID-19 Antigen Detection by the ASC ELISA Kit. Low levels of COVID-19 antigen were detected in human serum samples with the ASC ELISA kit.

Vendors Detection Limit
Applied StemCell

0.5 pg/ml

Competitor A

70 pg/ml

Competitor B

~1000 pg/ml

Competitor C

70 pg/ml

Competitor D 46.68 pg/ml
Competitor E

120 pg/ml

Competitor F 3.2 pg/ml
Competitor G

780 pg/ml

Competitor H

21.4 pg/ml

*Table 1: Low Detection Limits of Other Vendor ELISA kits. 

Case Study 2: Detection of VEGFA Antibodies for Therapeutics

Rabbit Line: Hu Rabbit

Reasons For Using the Rabbit Line: The antibodies produced by the Hu rabbit are more like human antibodies because they do not contain the disulfide bond found in most regular antibodies.

*Antibodies can be humanized successfully

Results: 11 bioactive mAbs detected; The 11 mAbs can be engineered into chimeric antibodies that hold human constant regions while maintaining high affinities (10x higher than Avastin)

Monoclonal Antibodies - VEGFA - Therapeutic Applications

Figure 2: Antibody Affinity. Bioactive, chimeric antibodies display higher affinities than Competitor I.

# mAbs expressed: 150

# high-affinity mAbs: 43

# Bioactive mAbs: 11

# Bioactive chimeric: 11

Table 2: Breakdown of Antibodies Expressed. Out of the 150 expressed mAbs, 43 were high-affinity antibodies. 11 bioactive antibodies were identified which can be engineered into chimeric antibodies.

Case Study 3: Identification of Fentanyl Antibodies For Drug Detection

Rabbit Line: HyImmune Rabbit

Reasons For Using the Rabbit Line:  Rapid production of small target antibodies

Results: Several fentanyl antibodies were discovered. 

*These antibodies were used to develop the ELISA kits. The ASC ELISA kit standard I-50 measurement was 10x more sensitive than other ELISA kits. The kit was even able to detect fentanyl derivatives.

Monoclonal Antibodies - Fentanyl Antibodies - Drug Selection

Figure 2: ASC ELISA Kit I-50 Measurements.   

How Our ELISA Kit Compares to Others

ASC ELISA kit

  • I-50 for fentanyl : 0.02 ng/ml
  • I-50 for carfentanil : 0.06 ng/ml

Competitor J ELISA kit

  • Fentanyl Kit : I-50 0.33 ng/ml
  • Carfentanil Kit : I-50 0.1 ng/ml

Table 3: How the ASC ELISA kit I-50 Measurements Compare to Other Kits. The ASC ELISA kit is 10x more sensitive.

Case Study 4: Customized Knockout Rabbit Antibody Generation

Rabbit Line: Gene Knockout Rabbit

Reasons For Using the Rabbit Line:  Knockout of the endogenous homolog protein generates an "all epitope targeting" environment for antibody production. This increases the rate of bioactive antibody identification. 

* We can generate multispecific antibodies for pre-clinical animal experiments

Results: Antibodies with high bioactivity were identified. These antibodies cross-react with both human and mouse targets.

Have Questions?

An Applied StemCell technical expert is happy to help, contact us today!

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