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人抗大肠杆菌蛋白IgG检测试剂盒
Human Anti-E.coli proteins IgG ELISA Kit
cat#500-110-ECP
INTENDED USE
The Human Anti-E. coli proteins IgG ELISA Kit is an indirectELISA suitable for detecting and quantifying antibody
activity(IgG) specific for E. coli (Escherichia coli) Host Cell Proteins(HCP) in serum, plasma or other
biological fluids. Antibodies mayoccur from either natural E. coli encounter or from
immunization with recombinant proteins expressed in E. coli. This kit is forresearch use only (RUO),
not for diagnostic purposes.
GENERAL INFORMATIONA
large number of genes have been cloned and expressed invarious host cells (E. coli, yeast,
baculovirus, NSO, Sp2/0, HEK,CHO cells). The translated recombinant proteins may remainwithin
the cell, requiring host cell disruption for release, and/ormay be secreted into the culture medium.
The targetrecombinant proteins (recProtein) would then be purified fromunwanted host cell protein
(HCP), often with the aid of a tag(e.g., His, GST, MBP). During the production of recProteins,
host cells die and decompose; thus, regardless of whether therecombinant product is obtained
from extracellular medium orafter disrupting the host cell, the entire repertoire of host cellproteins
present as potential contaminants in downstreampurification and processing of the recProtein
product.The presence of residual E. coli HCPs in purified recombinantproteins and its use as
antigens must be carefully studied beforeusing it for the detection of human antibodies.
SARS-NPexpressed in E. coli has resulted in false positive due to thedetection of E. coli antibodies
(refs 1). E. coli proteins (LPS,outer membrane proteins or residual HCPs) are highlyimmunogenic
in humans and animals. Most humans and animalshave exposure to E. coli and have varying levels
of circulatingantibodies. Therefore, human samples must be tested beforeand after exposure of
E. coli containing proteins or othermaterials.HCP in the processed recombinant may or may not
bedetectable by SDS-PAGE, ELISA, etc, which could have limitedsensitivity or specificity for the
particular E. coli protein(s)present. Immunization with preparations containing undetectedHCP,
however, often generate anti-HCP antibodies along withthe specific anti-recProtein antibodies.
Thus, the immunizationprocess can represent a more sensitive method than others fordisclosing
low level HCP contamination. Alternatively, theunwanted anti-HCP activity may obscure and/or
confuse theinterpretation of immunoassays designed to characterize andutilize the anti-recProtein
activity.
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