Premium
Isolation of glycoprotein D from herpes simplex virus type 1 by gel filtration high performance liquid chromatography
Author(s) -
McGarry Thomas J.,
AlAhdal Mohammed N.
Publication year - 1989
Publication title -
biomedical chromatography
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.4
H-Index - 65
eISSN - 1099-0801
pISSN - 0269-3879
DOI - 10.1002/bmc.1130030510
Subject(s) - chemistry , chromatography , glucosamine , size exclusion chromatography , monoclonal antibody , herpes simplex virus , glycoprotein , high performance liquid chromatography , methionine , affinity chromatography , monomer , biochemistry , antibody , virus , amino acid , virology , enzyme , biology , organic chemistry , immunology , polymer
Rabbit kidney (RK‐13) and human jejunum and ileum (I‐407) cells infected with herpes simplex virus type 1, strain F, were radiolabelled with [ 14 C]glucosamine or [ 35 S]methionine for 24 h. The cells were extracted with 1% Triton X‐100 and the extracts were separated by gel filtration high performance liquid chromatography. Monoclonal antibody immunoprecipitation of the fractions collected from the column revealed a monomeric glycoprotein D (gD) of 52 – 56 000 molecular weight from RK‐13 cells and two monomeric forms of gD, 54 000 and 58 000 molecular weight, from I‐407 cells. Densitometry scanning of the autoradiograms from SDS‐PAGE showed gD from the RK‐13 host cells to be 98.7% pure with the [ 35 S]methionine label and 97.0% pure with the [ 14 C]glucosamine. On the other hand, gD from the I‐407 host cells was only 78.6% with the [ 35 S]methionine label and 96% pure with the [ 14 C]glucosamine. This method could provide a means for the isolation of native gD for structural and immunological studies.