Simultaneous expression of displayed and secreted antibodies for antibody screen

PLoS One. 2013 Nov 11;8(11):e80005. doi: 10.1371/journal.pone.0080005. eCollection 2013.

Abstract

The display of full-length antibody on the cell surface was achieved by fusing a transmembrane domain of the platelet-derived growth factor receptor (PDGFR) to the C-terminus of the heavy chain constant region. We also incorporated a furin cleavage site between the constant region and PDGFR transmembrane domain to obtain secreted antibodies. As a result, antibodies can be expressed simultaneously on the cell surface in a membrane-anchored version for screening and selecting through fluorescence-activated cell sorting (FACS) analysis, as well as in conditioned medium in a secreted version for function analysis.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Antibodies / chemistry
  • Antibodies / genetics*
  • Antibodies / metabolism
  • CHO Cells
  • Cell Membrane / metabolism
  • Cricetulus
  • DNA Restriction Enzymes / genetics
  • DNA Restriction Enzymes / metabolism
  • Escherichia coli / genetics
  • Escherichia coli / metabolism
  • Furin / metabolism
  • Gene Expression
  • Genetic Vectors / chemistry*
  • HEK293 Cells
  • High-Throughput Screening Assays
  • Humans
  • Immunoglobulin Heavy Chains / chemistry
  • Immunoglobulin Heavy Chains / genetics*
  • Immunoglobulin Heavy Chains / metabolism
  • Protein Engineering
  • Proteolysis
  • Receptors, Platelet-Derived Growth Factor / genetics*
  • Receptors, Platelet-Derived Growth Factor / metabolism

Substances

  • Antibodies
  • Immunoglobulin Heavy Chains
  • Receptors, Platelet-Derived Growth Factor
  • DNA Restriction Enzymes
  • FURIN protein, human
  • Furin

Grant support

This work was partly supported by grants from the National Natural Science Foundation of China (#81173098 to SJ) and from Dgen Biotech Ltd., Hong Kong. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.