Scientist Solutions: International Life Science Community By Scientists For Scientists
Home » Protocols » Anti-idiotype Antibody Production
Thanks to our sponsors who make this site possible
Protocol for

Anti-idiotype Antibody Production

Would you like to save this topic, event, protocol or job so you can find it again easily?

Just click the "Save to My Lab Drawer" link and the item will be saved in the My Lab Drawer section of your bench space.

Available to members only. Please log in or register for your free account now.

Abstract or Description:

 Figure 1. Antibody idiotope— the unique set of antigenic determinants of the variable region of an antibody. The idiotype represents the variable part of an antibody, including the unique antigen binding site, and the combination of epitopes within the idiotype (i.e., the idiotopes) is unique for each antibody (Figure 1). When one antibody binds to one idiotope of another antibody it is referred to as an anti-idiotypic antibody (anti-ID). These highly specialized anti-idiotypic antibodies can be used in a PK assay to measure the drug level in patient samples and as a positive control or reference standard in any immune response (IR) assay (Figure 2).

Figure 2. Image shows anti-ID Fab antibody in purple, anti-ID full immunoglobulin in blue, drug target in red, drug in orange, anti-Fc antibody in gray, and HRP in orange. (A) PK Assay Format 1: Antigen Capture Assay; (B) PK Assay Format 2: Antigen Capture Assay with Complex-Specific Antibody; (C) PK Assay Format 3: Anti-Idiotypic Antibody Bridging Assay; (D) PK Assay Format 4: Anti-Idiotypic Antibody-Antigen Bridging Assay; (E) IR Assay: Anti-Idiotypic Antibody Bridging Assay.
Type 1. Anti-idiotypic antibody detecting FREE antibody. (paratope specific, inhibitory, neutralizing, detects free drug
Type 2. Anti-idiotypic antibody detecting TOTAL antibody. (not paratope specific, not inhibitory, detects total drug)
Type 3. Drug-target complex specific antibody detecting BOUND antibody. (not inhibitory, detects bound drug exclusively)
Type 4. Anti-idiotypic antibody detecting T cell receptor.
Three approaches in developing anti-Idiotypic Antibodies:
• Immunized phage displayed library technology: Target antibody in the forms of whole IgG, scFv, Fab or F(ab’)2 or peptides designed according to the sequences of the variable domains is utilized to immunize animals. Isolated immunized antibody library is packaged to phage library and anti-IDs can then be generated by biopanning for 3-4 cycles. scFv/Fab antibodies with an affinity of 10-10 M could be obtained. In vitro selection allows us to use 
isotype-matching control antibodies as blockers, to avoid enrichment of antibodies that bind to other regions of the target antibody
• Premade Non-Immunized human phage displayed library technology: phage display technology in the absence of immunization suits perfectly for the conformational epitope targeting anti-Idiotypic antibodies, since in vitro selection well maintains the correctly folded conformation of target antibody.
• Hybridomas technology: scFv/Fab forms of target antibodies are utilized to immunize rats or mice in the presence of isotype matching control antibodies for counter-selection.

Figures are showed on the related links. 

Attachments: No Attachments
Posted By: Candy Ton on 6/9/2015 8:54:56 AM
ChloeMica (Aug 19, 2015, 13:04 PM)
Great! Thanks for sharing. I'm interested in antibody stuff.
Add Comment