Anti-Belatacept Antibodies

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Antibodies for bioanalysis and drug monitoring of belatacept and biosimilars

• Develop highly selective and sensitive PK and ADA assays for belatacept (Nulojix) using our range of ready-made antibodies
• Recombinant antibodies specific to belatacept CTLA-4 domain
• Fully human format for surrogate positive control or calibrator
• Sequence defined, well characterized reagents with secure supply
• Stringent quality control for batch-to-batch consistency

Table 1. Antibodies Specific to Belatacept

* Affinity measured in the monovalent Fab format
¹ Fab-FH= Monovalent Fab antibody DYKDDDDK- and His-6-tags

Anti-Belatacept Antibodies

Belatacept is a genetically engineered fusion protein that consists of the functional binding domain of modified human cytotoxic T-lymphocyte antigen-4 (CTLA-4, CD152) and the Fc domain of human monoclonal immunoglobulin of the IgG1 subclass. The Fc part has the allotype IGHG1*05. Belatacept is only 2 amino acids different from abatacept (Orencia). The two amino acid substitutions (L104E and A29Y) give rise to slower dissociation rates for both CD86 and CD80. We offer the following antibodies:

• HCA391 (AbD37060, Fab-FH format) is a CTLA-4 domain-specific, inhibitory antibody that specifically recognizes the free human IgG1-Fc fusion protein belatacept and abatacept
• HCA392 (AbD37060ia, human IgG1 format) is a CTLA-4 domain-specific, inhibitory antibody that specifically recognizes the free human IgG1-Fc fusion proteins belatacept and abatacept
• HCA393 (AbD36935ia, human IgG1 format) is a CTLA-4 domain-specific, non-inhibitory antibody that specifically recognizes the free human IgG1-Fc fusion protein belatacept and abatacept

Human Anti-Belatacept Antibody, clone AbD37060 (HCA391), can be used for capture in a pharmacokinetic (PK) bridging ELISA to measure free drug, paired with Anti-Belatacept Antibody (HCA393) as a detection reagent. Human Anti-Belatacept Antibody, clone AbD37060ia (HCA392) and clone AbD36935ia (HCA393) are suitable as a surrogate positive control or calibrator for an anti-drug antibody (ADA) assay.

ELISA Protocols to Get You Started

PK Bridging ELISA

Fig. 1. Belatacept PK bridging ELISA using antibodies HCA391 and HCA393 .  

Schematic image of PK bridging ELISA measuring free drug. Capture antibody, Fab format (purple), fusion protein drug (green-gold), anti-drug detection antibody, Ig format (blue), labeled with HRP.

ADA Bridging ELISA Schematic for Belatacept

Fig. 2. Human anti-belatacept bridging ELISA for anti-drug antibody (ADA) assay development. A microtiter plate was coated over night with belatacept at a concentration of 1 µg/mL. After washing and blocking with PBST + 5% BSA, PBST with 10% human serum was added spiked with increasing concentrations of Human Anti-Belatacept clones AbD37060ia (HCA392) (black graph), AbD36935ia (HCA393) (blue graph) and AbD37065ia (HCA336) (red graph). Detection was performed using HRP-conjugated belatacept at a concentration of 2 µg/mL in HISPEC Assay Diluent followed by QuantaBlu™ Fluorogenic Peroxidase Substrate. Data are shown as the mean of three measurements. HRP-conjugation of belatacept was performed using a LYNX Rapid HRP Antibody Conjugation Kit®.

Schematic image of ADA bridging assay. Fusion protein drug as capture and detection reagent labeled with HRP (gold-green), monoclonal anti-drug antibody, Ig format (blue).

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Licensed Use: For in vitro research purposes and for commercial applications for the provision of in vitro testing services to support preclinical and clinical drug development. Any re-sale in any form or any other commercial application needs a written agreement with Bio-Rad.