Characterisation and application of antibodies specific for the long platelet-derived growth factor A and B chains
2004 (English)In: International Journal of Biochemistry and Cell Biology, ISSN 1357-2725, E-ISSN 1878-5875, Vol. 36, no 11, 2226-2241 p.Article in journal (Refereed) Published
The platelet-derived growth factor (PDGF) family comprises important mitogens for mesenchymal cells. The active dimeric form of PDGF consists of four structurally related A, B, C, and D chains. All PDGF-variants bind to PDGF-receptors. The A and B chains occur with and without basic C-terminal amino acid extensions as long (A(L) and B(L)) and short (A(S) and B(S)) isoforms. PDGF-A and -B form homo- or heterodimers. The biological relevance of short and long isoforms is unknown, although it may relate to different affinities for glycosaminoglycans of the cell glycocalix and intercellular matrix. Commercially available anti-PDGF-A and anti-PDGF-B antibodies cannot discriminate between the short and the long isoforms. Thus, to investigate the function of the long and short isoforms, we raised antibodies specific for the long A and B chain isoforms. The antibodies were affinity-purified and their properties analysed by surface plasmon resonance. Inhibition studies with different PDGF homodimers and dot-blot studies proved their high specificity for the respective isoforms. Both antibodies recognised the target PDGF homodimers complexed to the glycocalix of human arterial smooth muscle cells and human monocyte-derived macrophages. By using these specific antibodies, we were able to confirm at the protein level the synthesis of PDGF-A and -B during differentiation of human monocyte-derived macrophages and to demonstrate the presence of the PDGF-A(L) and PDGF-B(L) isoforms in human arterial tissue.
Place, publisher, year, edition, pages
Elsevier, 2004. Vol. 36, no 11, 2226-2241 p.
GAG, glycosaminoglycans, hASMC, hMDM, human arterial smooth muscle cells, human monocyte-derived macrophages, LMW, low molecular weight, mean fluorescence intensity, MFI, PDGF, PDGF isoforms, platelet-derived growth factor, resonance units, RU
Other Basic Medicine
Research subject Medical sciences
IdentifiersURN: urn:nbn:se:his:diva-11389DOI: 10.1016/j.biocel.2004.05.001ISI: 000223833700015PubMedID: 15313468ScopusID: 2-s2.0-4143146480OAI: oai:DiVA.org:his-11389DiVA: diva2:847241