EK-800-140-CHO 中国仓鼠卵巢细胞（CHO）宿主细胞蛋白（HCPS）ELISA检测试剂盒Chinese Hamster Ovary Cell (CHO) host cell Proteins (HCPs) ELISA kit, 96 tests
产品介绍：
A large number of genes have been cloned and expressed in various host cells (E. coli, yeast, baculovirus, NSO, Sp2/0, HEK, CHOcells). Recombinant proteins can either be secreted into the culture medium or remain inside the cells. The target recombinant proteins, with out without a tag (His, GST, MBP etc), must be purified to remove most of the unwanted, majority of the host cell’s protein (HCP). The traces of HCP present in the purified material do not represent a major problem for proteins that are used for in vitro or research use applications. However, many recombinant proteins are used for therapeutic purposes (Insulin, Erythropoietin, GM-CSF or humanized antibodies such Rituximab, Xolair rec), where the presence of HCP may be potentially toxic, allergic or create other health hazards. Therefore, it becomes necessary to determine the residual concentration of the HCP in the final recombinant material. Typically, ELISA and Western methods are used to determine concn of HCP in the samples. Western blot provides the nature (mol wt of the protein) of the contaminant and offer some idea about its concentration. This technique has limited used due to long assay, low sensitivity, and difficulty in processing large number of samples. ELISA, on the other hand, overcomes the limitations of the Western blot assay.
Since the processing the recombinant cells are different for each protein, it is difficult to predict what contaminant(s) will remain at the end of the process. It is possible that the major HCP are eliminated by certain purification protocol but other less abundant proteins may not. The detection method is based upon the use of antibodies. The most dominant proteins may not be immunogenic or less abundant may not be represented in the antigen mix. In addition, animals (goat, rabbit, or chicken) may make better antibodies to certain HCP protein. In short, the issue of residual HCP and the applicable detection methods are complex and they must be addressed specific to a given recombinant protein and the applicable purification protocol. We have used high avidity and high titer multi-host (Rabbit and Goat) anti-CHO HCP (secreted into the culture medium and the total cellular proteins) to offer the best recognition to CHO HCP. Antibodies have also been further characterized by SDS-PAGE, Western blot, and ELISA to detect both secreted and non-secreted proteins.
GSI’s CHO cell HCP ELISA kit is the most advanced, well characterized, and highly sensitive ELISA kit available in the market.
ELISA的基础是抗原或抗体的固相化及抗原或抗体的酶标记。结合在固相载体表面的抗原或抗体仍保持其免疫学活性，酶标记的抗原或抗体既保留其免疫学活性，又保留酶的活性。在测定时，受检标本（测定其中的抗体或抗原）与固相载体表面的抗原或抗体起反应。用洗涤的方法使固相载体上形成的抗原抗体复合物与液体中的其他物质分开。再加入酶标记的抗原或抗体，也通过反应而结合在固相载体上。此时固相上的酶量与标本中受检物质的量呈一定的比例。加入酶反应的底物后，底物被酶催化成为有色产物，产物的量与标本中受检物质的量直接相关，故可根据呈色的深浅进行定性或定量分析。