Preparation of high purity IgG and its application in screening for clones positive for Vibrio succinogenes L-asparaginase gene.
A previous paper reported application of L-asparaginase IgG from Vibrio succinogenes in screening for positive clones. This paper describes the detailed procedure for preparation of high-purity IgG, an essential step of which involves coupling of a cell-free extract of host E. coli Y 1090 to Sepharose 4B beads and adsorption of the non-specific antigen components by passage of the polyvalent IgG through this affinity column. The IgG thus obtained exhibited improved specificity and was utilized as a radioimmunologic and horseradish peroxidase-linked immunologic probe in screening for positive clones. These IgG probes had the advantages of low background, high sensitivity, and good reliability.
Guan
Y Q
YQ
Shanghai Institute of Plant Physiology, Academia Sinica.
Pong
H L
HL
He
Y S
YS
Chen
J M
JM
Ren
K Q
KQ
D001215:Asparaginase / Q000032:analysis / Q000235:genetics*; D015153:Blotting, Western; D003001:Cloning, Molecular; D004797:Enzyme-Linked Immunosorbent Assay; D015698:Genomic Library; D005779:Immunodiffusion; D007122:Immunoelectrophoresis; D007074:Immunoglobulin G / Q000276:immunology / Q000302:isolation & purification*; D014733:Vibrio / Q000201:enzymology / Q000235:genetics*
D007074:Immunoglobulin G; D001215:Asparaginase