Biotinylation of human C3.


Journal Article

Purified human C3 was biotinylated using the biotinyl-N-hydroxysuccinimide imidoester (BNHS). Depending on the input of BNHS, from three to six molecules of biotin were incorporated per C3 molecule. The biotinyl-C3 retained over 90% of its specific hemolytic activity and when bound to sheep erythrocytes maintained its ability to adhere to human C3b receptors. These functions could be blocked by avidin. The biotinyl-C3 was fragmented normally to C3c and C3d in human serum and adsorption with avidin-Sepharose indicated that biotin moities were present in both fragments. Fluorescein-conjugated avidin reacted well with cell-bound biotinyl-C3b and was useful for quantitating C3 fixation by flow cytometry. Ferritin-conjugated avidin was used as a marker to characterize the distribution of biotinyl-C3b on erythrocytes by electron microscopy. These results suggest that biotinyl-C3 and avidin derivatives may be very useful tools for studies of many of the biological functions of C3.

Full Text

Cited Authors

  • Berger, M; Gaither, TA; Cole, RM; Chused, TM; Hammer, CH; Frank, MM

Published Date

  • July 1982

Published In

Volume / Issue

  • 19 / 7

Start / End Page

  • 857 - 864

PubMed ID

  • 7121468

Pubmed Central ID

  • 7121468

International Standard Serial Number (ISSN)

  • 0161-5890

Digital Object Identifier (DOI)

  • 10.1016/0161-5890(82)90351-0


  • eng

Conference Location

  • England