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 with DYKDDDDK- and His-6-tags.
²Fab2-FH-X22-HRP = Bivalent Fab antibody with (F) DYKDDDDK, (H) His-6-tags, and Fab-SpyTag coupled to HRP-conjugated BiCatcher.

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
• TZA0144P (AbD36935pap) 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 Antibodies HCA393 or TZA0144P as a detection reagent. Human Anti-Belatacept Antibody clone AbD37060ia (HCA392), clone AbD36935ia (HCA393), and clone AbD36935pap (TZA0144P) 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.

Fig. 2. Belatacept PK bridging ELISA using antibodies HCA391 (capture) and TZA0144P (detection).    

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

ADA Bridging ELISA

Fig. 3. 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.