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First concept of antibody-based proteomics

A system for functional analysis of complementary DNA (cDNA)-encoded proteins was described in which selected portions of cDNAs are expressed as part of a fusion protein used for immunization to elicit antibodies. The concept of antibody-based proteomics was further developed and used in the HPA effort to generate more than 55,000 human recombinant proteins and 55,000 antibodies.

Key publication

  • Larsson M et al., A general bacterial expression system for functional analysis of cDNA-encoded proteins. Protein Expr Purif. (1996)
    PubMed: 8776765 DOI: 10.1006/prep.1996.0066

Other selected publications

  • Larsson M et al., High-throughput protein expression of cDNA products as a tool in functional genomics. J Biotechnol. (2000)
    PubMed: 10908795 DOI: 10.1016/s0168-1656(00)00258-3

  • Gräslund S et al., A high-stringency proteomics concept aimed for generation of antibodies specific for cDNA-encoded proteins. Biotechnol Appl Biochem. (2002)
    PubMed: 11916449 DOI: 10.1042/ba20010097

  • Gräslund S et al., A novel affinity gene fusion system allowing protein A-based recovery of non-immunoglobulin gene products. J Biotechnol. (2002)
    PubMed: 12204556 DOI: 10.1016/s0168-1656(02)00158-x

  • Falk R et al., An improved dual-expression concept, generating high-quality antibodies for proteomics research. Biotechnol Appl Biochem. (2003)
    PubMed: 12875650 DOI: 10.1042/BA20030091

  • Agaton C et al., Affinity proteomics for systematic protein profiling of chromosome 21 gene products in human tissues. Mol Cell Proteomics. (2003)
    PubMed: 12796447 DOI: 10.1074/mcp.M300022-MCP200

Figure legend: A flow chart representation of the basic high-stringency concept for the generation of antigen-specific antibodies to be used for antibody-based characterization of the human proteome. Adapted from Larsson et al. (2000). IMAC, immobilized metal affinity chromatography; His, histidine.

Key facts

  • The 1996 study outlined a new concept for systematic analysis of the human proteome
  • The recombinant proteins were used for both immunization and immunocapture
  • The principle was later used in the HPA program to generate 55,000 recombinant proteins
  • A search for “antibody-based proteomics” in Google Scholar yields more than 2,000 publications