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Origin of the Isoelectric Heterogeneity of Monoclonal Immunoglobulin h1B4 (1993)

Tsai, P. K. ; Bruner, Mark W. ; Irwin, Joseph I. ; [et al.]

Springer

Pharmaceutical research 10 (1993), S. 1580-1586

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ISSN: 1573-904X

Keywords: microheterogeneity ; monoclonal antibody ; deamidation ; isoelectric focusing

Source: Springer Online Journal Archives 1860-2000

Topics: Chemistry and Pharmacology

Notes: 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 pI 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 pI 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.

Type of Medium: Electronic Resource

URL: http://dx.doi.org/10.1023/A:1018912417607

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Microencapsulation selection for isolation of yeast mutants with increased secretion of Aspergillus awamori glucoamylase (1993)

Wittrup, K. D. ; Weber, A. L. ; Tsai, P.

New York, NY [u.a.] : Wiley-Blackwell

Biotechnology and Bioengineering 42 (1993), S. 351-356

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ISSN: 0006-3592

Keywords: microencapsulation ; selection ; secretion ; yeast ; Chemistry ; Biochemistry and Biotechnology

Source: Wiley InterScience Backfile Collection 1832-2000

Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology

Notes: We have developed a microencapsulation selection method which allows the rapid and quantitative screening of 〉106 yeast cells for enhanced secretion of Aspergillus awamori glucoamylase. The method provides a 400-fold single-pass enrichment for high-secreting mutants, and can be straightforwardly adapted for application to growth-based selection schemes with other microorganisms and enzymes. © 1993 John Wiley & Sons, Inc.

Additional Material: 2 Ill.