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High-resolution one-dimensional isoelectric focusing of mouse MHC class I antigens

Identification of natural and experimentally induced class I antigens

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Abstract

A method is described for a biochemical comparison of mouse class I antigens utilizing antisera with a monomorphic pattern of reaction and high-resolution one-dimensional isoelectric focusing (1D-IEF). The most commonly occurring and studied H-2K and D alleles were identified in a comparison of over 40 mouse strains. By comparing H-2 mutant mouse strains, cell lines transfected with defined class I genes, or mice transgenic for a mouse class I gene and H-2 recombinant mouse strains, unambiguous identification of class I alleles was possible. The complex pattern presented by H-2-heterozygous mice was readily resolved into the contribution by the individual parental alleles. The H-2 b bm series of mutants was analyzed, and for those mutants where a charge difference was predicted based on their known sequence, a change in isoelectric point (IEP) was indeed observed. Based on analysis by IEF, the bm8 mutant may contain (an) amino acid substitution(s) in addition to those reported. The present method further appears useful in elucidating defects in class I antigen synthesis and post-translational modifications, as these cannot be easily characterized when using surface staining with monoclonal antibodies (mAbs).

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Authors and Affiliations

  1. Department of Tumor Biology, Karolinska Institutet, Box 60 400, S-104 01, Stolkholm, Sweden

    Hans-Gustaf Ljunggren & Maria G. Masucci

  2. Department of Cellular Biochemistry, The Netherlands Cancer Institute, Antoni van Leeuwenhoek Huis, Plesmanlaan 121, 1066 CX, Amsterdam, The Netherlands

    Hidde L. Ploegh

  3. Department of Molecular Genetics, The Netherlands Cancer Institute, Antoni van Leeuwenhoek Huis, Plesmanlaan 121, 1066 CX, Amsterdam, The Netherlands

    Margriet Oudshoorn-Snoek

Authors
  1. Hans-Gustaf Ljunggren
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  2. Margriet Oudshoorn-Snoek
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  3. Maria G. Masucci
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  4. Hidde L. Ploegh
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Ljunggren, HG., Oudshoorn-Snoek, M., Masucci, M.G. et al. High-resolution one-dimensional isoelectric focusing of mouse MHC class I antigens. Immunogenetics 32, 440–450 (1990). https://doi.org/10.1007/BF00241639

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  • DOI: https://doi.org/10.1007/BF00241639

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