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Monoclonal antibodies against Galα 1 −4Galß 1 −4Glc(P k , CD77) produced with a synthetic glycoconjugate as immunogen: Reactivity with carbohydrates, with fresh frozen human tissues and hematopoietic tumors
Author(s) -
Oosterwlik Egbert,
Kalisiak Angela,
Wakka Jeannette C.,
Scheinberg David A.,
Old Lloyd J.
Publication year - 1991
Publication title -
international journal of cancer
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 2.475
H-Index - 234
eISSN - 1097-0215
pISSN - 0020-7136
DOI - 10.1002/ijc.2910480610
Subject(s) - immunogen , monoclonal antibody , glycoconjugate , microbiology and biotechnology , epitope , biology , glycolipid , staining , monoclonal , antibody , biochemistry , immunology , genetics
The purpose of this study was to isolate mouse monoclonal antibodies (MAbs) recognizing Galα I ‐4Galβ I −4Glc (P k . CD77), which has been described as a Burkitt's‐lym homa‐associated using a synthetic glycoconjugate as immunogen and a filter immunoplaque screening assay. One MAb (PK67) was characterized by immuno‐thin‐layer chromatography, ELISA and competition assays using neutral glycolipids from various sources and a variety of carbohydrates and glycoproteins. Epitope analysis showed that all 3 carbohydrate moieties are required in PK67 binding. No cross‐reactivity was observed with closely related carbohydrate structures, with the exception of Galα I ‐4Ga1β I ‐4GlcNAc. However, the reactivity with this structure was several orders of magnitude lower than with P k . lmmunohistochemical analysis of fresh‐frozen tissue specimens showed that Gb 3 , is widely expressed: prominent staining was observed in many normal tissues, e.g., kidney and gastric tissue and on capillary endothelial cells. In lymph nodes, very weak staining of a few B cells was observed. Flow cytometric analysis of hematopoietic neoplasms showed Gb, expression on B‐cell neoplasms, particularly those with committed 8‐cell phenotype indicating that, in addition to earlier reports of Gb 3 , expression on Burkitt's lymphoma, Gb 3 , is present on other committed B‐cell neoplasms.