人抗大腸桿菌蛋白IgG檢測 Anti E.coli proteins IgG ELISA Kit 說明書

人抗大腸桿菌蛋白IgG檢測試劑盒

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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