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This bispecific is comprised of single-chain variable fragments (scFv) of two antibodies. An scFv is a recombinant antibody construct that’s made up of the variable regions of an antibody’s heavy (VH) and light (VL) chains, connected by a flexible peptide linker. The linker is usually 10–25 amino acids long. scFvs are about 25–30 kDa in size and retain the antigen-binding properties of the corresponding IgG antibody.
This particular construct has a molecular weight of 54kDa in a single, covalently linked molecule. Since each arm has a stripped-down binding site, the affinity for their targets is weaker than in the parent antibodies. The Kd dissociation constant of the one arm to the CD3 antigen receptor on T cells is 2.6 x 10-7 M, and the Kd of the other arm to CD19 surface antigen on B lymphoma cells (and normal B lymphocytes) has a Kd of 1.49 x 10-9 M. typically high binding ELISA antibodies have strong affinities with Kd values of 10‑-9 to 10-11 M.
A typical sandwich ELISA utilizes a coat antibody to one epitope on the analyte to be measured, and a second antibody to a second epitope on the analyte. In this case, the coat reagent is the target of the bispecific construct, CD3 protein, and the detection reagent is the other protein, CD19. Therefore, binding of the bispecific to the CD3 protein coat is especially weak and tends to fall off during washing steps.
The method, shown in outline below, achieves a sensitivity of 15pg/mL in buffer and 60pg/mL in neat serum (15pg/mL in 25% serum).
MarinBio has validated this bispecific antibody ELISA to FDA GLP standards for PK analysis, and can adapt the technology to detect other bispecific antibodies in GLP or cGMP assays. Click for details:https://www.marinbio.com/novel-high-sensitivity-bispecific-antibody-elisa/
Contact us to discuss the specifics of your project. Our team of experienced scientists is ready to assist you with tailored solutions to meet your unique needs. Whether you need help with assay development, FDA IND, stability study, lot release approval, or other scientific endeavors, we are here to support you.
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Yes, we have over 30 years of expertise in developing and validating cell based assays for FDA IND, lot release or stability study approval. Our team can also liaise directly with the FDA on your behalf, ensuring a smooth and compliant process. Please contact us to learn more about how we can assist you with your assay development and FDA approval needs.
MarinBio scientists are experienced and creative in designing, developing, validating assays. Some of the assays we perform are listed below, but there are many more that we created, developed and validated. Give us call so that we may help you.
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With 30 years of expertise, we specialize in developing ELISAs essential for potency assay, lot release tests, IND/BLA applications, and clinical trials. Our cGMP/GLP services offer:
MarinBio’s scientists have developed ELISAs to detect analytes in complex matrices like blood, plasma, serum, and cell culture media. They created a 90-minute GMP ELISA, manufactured thousands of kits, and distributed them to hospital med-tech labs, training technicians for clinical treatment differentiation.
We maintain direct interactions with the FDA, both independently and through our clients.
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