Origin of the Isoelectric Heterogeneity of Monoclonal Immunoglobulin h1B4

P. K. Tsai, Mark W. Bruner, Joseph I. Irwin, Charlotte C Yu Ip, Cynthia N. Oliver, Randall W. Nelson, David B. Volkin, C. Russell Middaugh

Research output: Contribution to journalArticle

50 Citations (Scopus)

Abstract

The origin of the microheterogeneity of a highly purified antiinflammatory humanized monoclonal antibody prepared in mammalian cell culture has been investigated. This antibody is an IgG directed toward human CD 18 (a subunit of leukocyte integrins). When the IgG preparation is subjected to isoelectric focusing, it is found to contain four major species with p I values ranging from 6 to 7. Although the relative amounts of each form differ and some species are present only in small quantities, each has been isolated by a combination of high-resolution anion-exchange chromatography and isoelectric focusing. Comparative studies reveal no detectable differences in overall secondary (far UV circular dichroism) or tertiary (intrinsic fluorescence) structure, molecular weight (laser-desorption mass spectroscopy), or antigen binding activity. When each of the isolated species is incubated under conditions which favor deamidation, it is converted to forms of lower p I which appear to correspond to naturally observed species. While the isolated light chain is relatively homogeneous, the heavy chain exhibits a pattern of isoelectric focusing bands similar to that of the intact immunoglobulin. These results suggest that in this case, charge microheterogeneity is due to the sequential deamidation of the immunoglobulin heavy chain.

Original languageEnglish (US)
Pages (from-to)1580-1586
Number of pages7
JournalPharmaceutical Research
Volume10
Issue number11
DOIs
StatePublished - 1993
Externally publishedYes

Fingerprint

Isoelectric Focusing
Immunoglobulins
Immunoglobulin G
Antibodies, Monoclonal, Humanized
Immunoglobulin Heavy Chains
Dichroism
Circular Dichroism
Chromatography
Cell culture
Integrins
Anions
Desorption
Mass Spectrometry
Lasers
Leukocytes
Anti-Inflammatory Agents
Cell Culture Techniques
Molecular Weight
Fluorescence
Molecular weight

Keywords

  • deamidation
  • isoelectric focusing
  • microheterogeneity
  • monoclonal antibody

ASJC Scopus subject areas

  • Biotechnology
  • Molecular Medicine
  • Organic Chemistry
  • Pharmacology (medical)
  • Pharmaceutical Science
  • Pharmacology
  • Chemistry(all)

Cite this

Tsai, P. K., Bruner, M. W., Irwin, J. I., Ip, C. C. Y., Oliver, C. N., Nelson, R. W., ... Middaugh, C. R. (1993). Origin of the Isoelectric Heterogeneity of Monoclonal Immunoglobulin h1B4. Pharmaceutical Research, 10(11), 1580-1586. https://doi.org/10.1023/A:1018912417607

Origin of the Isoelectric Heterogeneity of Monoclonal Immunoglobulin h1B4. / Tsai, P. K.; Bruner, Mark W.; Irwin, Joseph I.; Ip, Charlotte C Yu; Oliver, Cynthia N.; Nelson, Randall W.; Volkin, David B.; Middaugh, C. Russell.

In: Pharmaceutical Research, Vol. 10, No. 11, 1993, p. 1580-1586.

Research output: Contribution to journalArticle

Tsai, PK, Bruner, MW, Irwin, JI, Ip, CCY, Oliver, CN, Nelson, RW, Volkin, DB & Middaugh, CR 1993, 'Origin of the Isoelectric Heterogeneity of Monoclonal Immunoglobulin h1B4', Pharmaceutical Research, vol. 10, no. 11, pp. 1580-1586. https://doi.org/10.1023/A:1018912417607
Tsai PK, Bruner MW, Irwin JI, Ip CCY, Oliver CN, Nelson RW et al. Origin of the Isoelectric Heterogeneity of Monoclonal Immunoglobulin h1B4. Pharmaceutical Research. 1993;10(11):1580-1586. https://doi.org/10.1023/A:1018912417607
Tsai, P. K. ; Bruner, Mark W. ; Irwin, Joseph I. ; Ip, Charlotte C Yu ; Oliver, Cynthia N. ; Nelson, Randall W. ; Volkin, David B. ; Middaugh, C. Russell. / Origin of the Isoelectric Heterogeneity of Monoclonal Immunoglobulin h1B4. In: Pharmaceutical Research. 1993 ; Vol. 10, No. 11. pp. 1580-1586.
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