A novel VHH antibody targeting the B cell-activating factor for B-cell lymphoma

Int J Mol Sci. 2014 May 28;15(6):9481-96. doi: 10.3390/ijms15069481.

Abstract

Objective: To construct an immune alpaca phage display library, in order to obtain a single domain anti-BAFF (B cell-activating factor) antibody.

Methods: Using phage display technology, we constructed an immune alpaca phage display library, selected anti-BAFF single domain antibodies (sdAbs), cloned three anti-BAFF single-domain antibody genes into expression vector pSJF2, and expressed them efficiently in Escherichia coli. The affinity of different anti-BAFF sdAbs were measured by Bio layer interferometry. The in vitro biological function of three sdAbs was investigated by cell counting kit-8 (CCK-8) assay and a competitive enzyme-linked immunosorbent assay (ELISA).

Results: We obtained three anti-BAFF single domain antibodies (anti-BAFF64, anti-BAFF52 and anti-BAFFG3), which were produced in high yield in Escherichia coli and inhibited tumor cell proliferation in vitro.

Conclusion: The selected anti-BAFF antibodies could be candidates for B-cell lymphoma therapies.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Animals
  • B-Cell Activating Factor / immunology*
  • B-Lymphocytes / drug effects
  • B-Lymphocytes / immunology
  • B-Lymphocytes / pathology
  • Camelids, New World
  • Cell Line, Tumor
  • Cell Proliferation / drug effects
  • Cloning, Molecular
  • Escherichia coli / genetics
  • Humans
  • Immunoglobulin Heavy Chains / chemistry
  • Immunoglobulin Heavy Chains / genetics
  • Immunoglobulin Heavy Chains / immunology
  • Immunoglobulin Heavy Chains / pharmacology
  • Lymphoma, B-Cell / drug therapy*
  • Lymphoma, B-Cell / immunology
  • Molecular Sequence Data
  • Peptide Library*
  • Sequence Alignment
  • Single-Domain Antibodies / chemistry
  • Single-Domain Antibodies / genetics
  • Single-Domain Antibodies / immunology*
  • Single-Domain Antibodies / pharmacology*

Substances

  • B-Cell Activating Factor
  • Immunoglobulin Heavy Chains
  • Peptide Library
  • Single-Domain Antibodies