Identification of a functional domain of human granulocyte colony-stimulating factor using neutralizing monoclonal antibodies
| Autor: | Je, Layton, Morstyn G, louis fabri, Ge, Reid, Aw, Burgess, Rj, Simpson, Ec, Nice |
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| Rok vydání: | 1991 |
| Předmět: |
Models
Molecular Protein Conformation Molecular Sequence Data Antibodies Monoclonal Enzyme-Linked Immunosorbent Assay Peptide Fragments Recombinant Proteins Cell Line Epitopes Neutralization Tests Endopeptidases Granulocyte Colony-Stimulating Factor Humans Amino Acid Sequence Binding Sites Antibody Cell Division Chromatography High Pressure Liquid Software |
| Zdroj: | Europe PubMed Central |
| ISSN: | 0021-9258 |
| Popis: | Human granulocyte colony-stimulating factor (G-CSF) is a hemopoietic growth factor that is being used successfully to treat various forms of neutropenia. To define functionally important regions of G-CSF, we have prepared 37 monoclonal anti-G-CSF antibodies and mapped the regions of G-CSF recognized by different antibody groups. Antibodies recognizing similar epitopes were identified by competition assays, neutralization assays, conformation dependence and cross-reactivity with canine G-CSF. Seven of eight neutralizing antibodies fell into two related epitope groups and were conformation-dependent. The eighth was unrelated and conformation-independent. Peptides of G-CSF were generated by chemical or enzymatic digestion and tested for antibody reactivity. One of the neutralizing antibodies (LMM351) recognized a small, disulfide-bonded peptide from the V8 protease digest (residues 34-46). A synthetic peptide (residues 20-58) was recognized by all the neutralizing antibodies, implicating this disulfide-bonded loop in receptor binding. The epitopes recognized by nonneutralizing antibodies were found throughout G-CSF. Thus, regions of G-CSF that are not involved in receptor binding have also been defined. A CNBr peptide (residues 1-121) had greatly reduced biological activity, indicating that the COOH terminus is required for receptor binding. We predict that residues 20-46 and the COOH terminus bind to the G-CSF receptor. |
| Databáze: | OpenAIRE |
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