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  • 1
    Electronic Resource
    Electronic Resource
    Marsupial light chains: IGK with four V families in the opossum Monodelphis domestica (1999)
    Springer
    Immunogenetics 50 (1999), S. 329-335 
    Details
    ISSN: 1432-1211
    Keywords: Key words Marsupials ; Light chains ; Variable regions ; IGK ; Evolution
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract  A full-length and several partial cDNAs encoding IGK light chains from the marsupial South American opossum, Monodelphis domestica, were isolated and characterized. Using these clones as a starting point, the expressed IGKV repertoire was sampled by anchored polymerase chain reaction using an IGKC-specific primer. Based on nucleotide sequences of twenty unique, expressed IGKV-J combinations, there are at least four IGKV families and two J segments. Southern blot analysis revealed each IGK-V family contains multiple gene segments totaling at least thirty-five IGKV in the opossum genome. No evidence for particular, recurrent IGKV-J combinations in the opossum IGK repertoire was seen, rather the V-J combinations appeared random and diverse. Each of the four IGKV families appear more closely related to V segments from placental mammals than to each other, suggesting the duplication of the IGKV families prior to the separation of marsupials and placental mammals more than one-hundred-million years ago. Overall, the complexity of opossum light chain V segments appears greater than that found in the heavy chain, and light chains are likely to contribute significantly to Ig diversity in this species.With this report, the homologues encoding all three classes of eutherian Ig chains, IGH, IGL, and IGK, have been described in a non-placental mammal.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/s002510050609
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    Paper (German National Licenses)
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  • 2
    Electronic Resource
    Electronic Resource
    The glycosylation pattern of a humanized IgGl antibody (D1.3) expressed in CHO cells (1997)
    Springer
    Glycoconjugate journal 14 (1997), S. 201-207 
    Details
    ISSN: 1573-4986
    Keywords: humanized IgG antibody ; immunotherapy ; recombinant glycoprotein ; CHO cell glycosylation ; HPAEC
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Abstract A humanized IgG antibody (D1.3) which retains murine complementarity determining regions specific for the antigen lysozyme has been expressed in CHO-DUKX cells. Heavy and light chain containing plasmids were co-transfected into CHO-DUKX cells and stable clones were grown in DMEM/F12 medium supplemented with 5% foetal calf serum. D1.3 antibody was purified from culture supernatants by Protein G chromatography. With the recombinant D1.3 antibody as a model, this cell culture system was shown to glycosylate the IgG Fc region in a similar manner to IgG isolated from serum. The neutral, core fucosylated biantennary oligosaccharides found are present in serum IgG and no novel carbohydrate sequences were detected. The degree of terminal agalactosylation was also similar to normal serum, in contrast to the increased levels found in rheumatoid serum. Furthermore, those oligosaccharides which lack only one terminal Gal are exclusively galactosylated on the GlcNAc(β1,2) Man(α1,6) Man(β1,4) antenna. Unambiguous identification of the exact glycosylation pattern of the antibody was carried out by a combination of specific exoglycosidase digestions, gel permeation chromatography of 2-aminobenzamide derivatives, high pH anion exchange chromatography and methylation analysis followed by gas–liquid chromatography-mass spectrometry. Abbreviations: CDR, complementarity determining region; CHO, chinese hamster ovary; GPC, gel permeation chromatography; 2-AB, 2-aminobenzamide; HPAEC-PAD, high pH anion exchange chromatography with pulsed amperometric detection; GC-MS, gas chromatography with mass spectrometry analysis; PNGase F, peptide-N-glycosidase F; PGC, porous graphitized carbon column; RAAM, reagent array analysis method; NeuAc: N-acetylneuraminic acid; NeuGc: N-glycolylneuraminic acid
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1023/A:1018589704981
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  • 3
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    Marsupial light chains: IGK with four V families in the opossum Monodelphis domestica (1999)
    Springer
    Details
    Publication Date: 1999-12-02
    Print ISSN: 0093-7711
    Electronic ISSN: 1432-1211
    Topics: Biology , Medicine
    Published by Springer
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