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3 Rabbit monoclonal antibody (Mab) is secreted by the plasma B-cell of the rabbit. Traditional generation of rabbit Mab relies on a rabbit myeloma for B- cell fusion (<1% fusion rate) and hybridoma development. ARM uses a non-fusion Fab antibody library to produce rabbit Mab in vitro. ARM platform utilizes the entire rabbit antibody repertoire to screen for high affinity and high specificity Fab binders with broad epitope coverage to antigen of interest. ARM has higher antibody screening throughput and success rate. ARM Fab is amenable to antibody engineering including F(ab)2, IgG, scfv and multitude of different Fc and non-fc conjugates. ARM technology is the solution to high quality reagents for research and commercial developments.
4 3 months 1.5 months 1.5 months 1 month
5 Antigen : 1.5 mg protein/rabbit (or 3mg KLH conjugated peptide/rabbit) Adjuvant : CFA/IFA Rabbit : New Zealand White (NZW) Duration : 2-3 months QC Data : Serum ELISA Serum ELISA Serum is QC at seven dilutions: 1562x, 3125x, 6250x, 12500x, 25000x, 50000x, x, x), PASS condition: OD at the X diluted serum> bg x 2 with coating 0.2ug immunogen
6 Plasmid Vector : Phagemid Phage : Helper Phage Library size : 10 8 or higher Host Cell : E. coli QC Data : Fab Antibody PCR Fab Antibody Diversity Sequencing
7 Screening Antigen : 350ug protein/library or 1mg BSA conjugated peptide + 1mg free form peptide/rabbit Induction Host cell : E. coli QC Data : Fab Antibody ELISA Fab Antibody Sequencing
8 Plasmid Vector : pab03(with Fc) + Fd pab04 + LC Host Cell : 293H QC Data : IgG Antibody Coomassie Blue IgG Antibody ELISA A A A Protein A purification
9 Comparison of Anti-TSH Rabbit Mab With a Commercial Antibody TSH Dose Respnse Curves Comparison of isolated rabbit Mab for TSH (thyroid stimulating hormone) with a commercial antibody diagnostic kit on detection of TSH in a sandwich ELISA setting. The rabbit Mab shows a superior binding activity to TSH compared side-byside to the commercial antibody. Validation of Isolated Anti-Collagen Rabbit Mab by Sandwich ELISA NTx Standard Curve A rabbit Mab was used to capture a collagen peptide (NTx) with serial dilutions (0, 0.05, 0.10, 0.25, 0.50 and 1.00 ug/ml) in physiologic relevant concentration range in a sandwich ELISA setting.
10 Technology Abnova Rabbit Mab (ARM ) Non-Fusion Fab Antibody Library Conventional Rabbit Mab Rabbit Myeloma Antibody Repertoire Broad (no fusion needed) Narrow (<1% fusion rate) Screening Throughput High Low Epitope Coverage High Low Antibody Success Rate Higher Lower Antibody Yield High Low Clone Stability Excellent Poor Antibody Engineering Readily Applicable Troublesome
11 Rabbit Mouse Immune Tolerance None Yes Antibody Diversity Broad Narrow Antigenicity Strong reaction to small molecules, peptides, & proteins. Weak reaction to small molecules and peptides. Affinity Picomolar Nanomolar Specificity High Moderate Isotype IgG IgG1, IgG2a, IgG2b, IgG3 Applications Especially suited for IHC, IP, FC, antibody pair & kit. Antibody Engineering Yes Yes Mostly suited for WB, ELISA, IF, antibody pair & kit.
12 F(ab)2 + different species Fc (rabbit, mouse, rat, human) Fab F(ab)2 F(ab)2 + different isotypes Fc (IgG1, IgG2a, IgG2b, IgG3, IgG4) F(ab)2 + Conjugates (Biotin, Streptavidin, HRP)
13 What is the difference between Abnova and Epitomics technology? Abnova uses non-fusion antibody library derived from entire immune repertoire instead of myeloma fusion (<1% fusion rate) for Mab generation and screening. Why choose rabbit Mab over mouse Mab? Rabbit Mab has broader antibody diversity, antigenicity, affinity, specificity, and applicability than mouse Mab. How many unique antibody clones will be generated? It depends on the size (e.g., peptide vs. protein) and sequence homology (e.g., human vs. rabbit) of the antigen. Who owns the sales and IP right of the rabbit Mab? Customer How much is charged if antibody production is not successful? Antibody production charge is based on milestones, hence minimizes the risk to the customer. Can rabbit Mab recognize hapten or phospho-peptide? Yes.
14 Can rabbit Fab antibody bind to Protein G? Yes. Can F(ab)2 antibody be conjugated to other tag proteins? Yes, including different species Fc, isotype Fc, biotin, streptavidin, HRP. Can customer access the plasmid cdna of rabbit Mab? No. The plasmids are Abnova s proprietary technology. Can customer access the antibody sequence of rabbit Mab? Yes. There is an extra charge for the antibody sequence but it can be decreased or waived depending on the scale of follow-up recombinant antibody production. Are adjunctive services (IHC, antibody pairing & scaled-up) available to customer? Yes.
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