Fractionation of the human plasma proteome for monoclonal antibody proteomics-based biomarker discovery 2: Antigen identification by dot-blot array screening

András Kovács, Zoltán Patai, A. Guttman, János Kádas, László Takács, István Kurucz

Research output: Contribution to journalArticle

1 Citation (Scopus)

Abstract

Immunization with complex mixtures, like the human plasma resulted in the generation of cloned mAb libraries (PlasmaScan™ and QuantiPlasma™ libraries, with >1000 individual mAbs) reacting with a nonredundant set of antigenic epitopes. mAb proteomics refers to quasi-hypothesis-free profiling of plasma samples with nascent or cloned mAb libraries for the discovery of disease-specific biomarkers. Once mAbs with biomarker potential have been identified, the next task is the determination of cognate antigens recognized by the respective mAbs. To determine the cognate protein antigen corresponding to each individual mAbs in the cloned mAb libraries, we have separated human plasma by consecutive steps of desalting and various chromatography procedures. The process resulted in 783 fractions, which we termed "Analyte Library" (AL). The AL represents the human plasma proteome in relatively low-protein complexity fractions. Here, to determine the utility of the AL, we selected ten plasma proteins and checked for their presence in the fractions. Among the ten cases, the distribution of four selected plasma proteins matched expectations, as these proteins were present only in a few fractions corresponding to their physical, chemical, and biochemical properties. However, in six cases, we observed "smear" -like distribution or complete absence of the proteins, suggesting that protein-protein interactions or protein variants may alter the observed plasma distribution profiles. Nevertheless, we conclude that the AL is an efficient, high throughput tool to complement the mAb biomarker discovery process with cognate protein antigen identification for each mAbs.

Original languageEnglish
Pages (from-to)3064-3071
Number of pages8
JournalElectrophoresis
Volume34
Issue number20-21
DOIs
Publication statusPublished - Nov 2013

Fingerprint

Plasma (human)
Biomarkers
Proteome
Fractionation
Proteomics
Libraries
Screening
Monoclonal Antibodies
Antigens
Proteins
Blood Proteins
Immunization
Plasmas
Salt removal
Beam plasma interactions
Chromatography
Complex Mixtures
Epitopes
Throughput

Keywords

  • Analyte Library
  • Biomarkers
  • Dot blot
  • MAb proteomics
  • Mass spectrometry

ASJC Scopus subject areas

  • Biochemistry
  • Clinical Biochemistry

Cite this

Fractionation of the human plasma proteome for monoclonal antibody proteomics-based biomarker discovery 2 : Antigen identification by dot-blot array screening. / Kovács, András; Patai, Zoltán; Guttman, A.; Kádas, János; Takács, László; Kurucz, István.

In: Electrophoresis, Vol. 34, No. 20-21, 11.2013, p. 3064-3071.

Research output: Contribution to journalArticle

Kovács, András ; Patai, Zoltán ; Guttman, A. ; Kádas, János ; Takács, László ; Kurucz, István. / Fractionation of the human plasma proteome for monoclonal antibody proteomics-based biomarker discovery 2 : Antigen identification by dot-blot array screening. In: Electrophoresis. 2013 ; Vol. 34, No. 20-21. pp. 3064-3071.
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