Novel purification method of human immunoglobulin by using a thermo-responsive protein A

Ichiro Koguma, Shuntaro Yamashita, Satoshi Sato, Kazuo Okuyama, Yoshinori Katakura

Research output: Contribution to journalArticle

9 Citations (Scopus)

Abstract

We attempted to evaluate a novel purification method of immunoglobulins (IgGs) by using a mutant type of protein A. Although this mutant protein A binds to IgGs at 5 °C, the IgGs are released at 40 °C; hence, it was designated as thermo-responsive protein A (TRPA). We aimed to purify IgG1 from the culture supernatant of CHO cells producing AE6F4 human monoclonal IgG1. AE6F4 IgG1 was purified using only a TRPA-filled column and by modifying the temperature, without any exposure to acidic conditions. Furthermore, the purified AE6F4 IgG1 maintained the inherent binding affinity to antigen, while this property was lost in AE6F4 IgG1 purified using a conventional protein A (CPA) column possibly because of product aggregation and fragmentation. These data suggest that IgG is sensitive to acid treatment; however, it can be highly purified with retention of high affinity by using a TRPA column. Further, this purification method can be used on an industrial scale for the purification of antibody drugs.

Original languageEnglish
Pages (from-to)149-153
Number of pages5
JournalJournal of Chromatography A
Volume1305
DOIs
Publication statusPublished - Aug 30 2013

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Staphylococcal Protein A
Purification
Immunoglobulins
Immunoglobulin G
Mutant Proteins
CHO Cells
Distillation columns
Agglomeration
Antigens
Temperature
Acids
Antibodies
Pharmaceutical Preparations

All Science Journal Classification (ASJC) codes

  • Analytical Chemistry
  • Biochemistry
  • Organic Chemistry

Cite this

Novel purification method of human immunoglobulin by using a thermo-responsive protein A. / Koguma, Ichiro; Yamashita, Shuntaro; Sato, Satoshi; Okuyama, Kazuo; Katakura, Yoshinori.

In: Journal of Chromatography A, Vol. 1305, 30.08.2013, p. 149-153.

Research output: Contribution to journalArticle

Koguma, Ichiro ; Yamashita, Shuntaro ; Sato, Satoshi ; Okuyama, Kazuo ; Katakura, Yoshinori. / Novel purification method of human immunoglobulin by using a thermo-responsive protein A. In: Journal of Chromatography A. 2013 ; Vol. 1305. pp. 149-153.
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