A comparative study of capillary electrophoresis and isothermal titration calorimetry for the determination of binding constant of human serum albumin to monoclonal antibody

Melinda Andrási, Gábor Lehoczki, Zoltán Nagy, G. Gyémánt, András Pungor, A. Gáspár

Research output: Contribution to journalArticle

11 Citations (Scopus)

Abstract

This paper focuses on the investigation of the interactions between the anti-HSA-mAb and its protein antigen using CZE, ACE, and isothermal titration calorimetry. The CZE revealed the formation of the anti-HSA-mAb·HSA and anti-HSA-mAb·(HSA)2 complexes and the binding constants determined by plotting the amount of the bound anti-HSA-mAb as a function of the concentration of HSA. The ACE provided information on the binding strength from the change in effective electrophoretic mobility of the anti-HSA-mAb. These two separation techniques estimated the presence of two binding sites. The equilibrium dissociation constant values obtained by CZE and ACE were found to be 2.26 × 10-6 M for anti-HSA-mAb·HSA, 1.22 × 10-6 M for anti-HSA-mAb·(HSA)2 and 4.45 × 10-8 M for anti-HSA-mAb·HSA, 1.08 × 10-7 M for anti-HSA-mAb·(HSA)2, respectively. The dissociation constant data obtained by ACE were in congruence with the values obtained by isothermal titration calorimetry (2.74 × 10-8 M, 1.04 × 10-7 M).

Original languageEnglish
Pages (from-to)1274-1281
Number of pages8
JournalElectrophoresis
Volume36
Issue number11-12
DOIs
Publication statusPublished - Jun 1 2015

Fingerprint

Capillary electrophoresis
Calorimetry
Capillary Electrophoresis
Titration
Serum Albumin
Monoclonal Antibodies
Electrophoretic mobility
Binding Sites
Antigens
Proteins

Keywords

  • ACE
  • Binding constant
  • CZE
  • HSA
  • MAb

ASJC Scopus subject areas

  • Biochemistry
  • Clinical Biochemistry

Cite this

A comparative study of capillary electrophoresis and isothermal titration calorimetry for the determination of binding constant of human serum albumin to monoclonal antibody. / Andrási, Melinda; Lehoczki, Gábor; Nagy, Zoltán; Gyémánt, G.; Pungor, András; Gáspár, A.

In: Electrophoresis, Vol. 36, No. 11-12, 01.06.2015, p. 1274-1281.

Research output: Contribution to journalArticle

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