bordsrening och ÄKTA rening för den nativa antikroppsreningen. Proteinsckvens An

Question

bordsrening och ÄKTA rening för den nativa antikroppsreningen. Proteinsckvens Anti-HER2 Light chain 1 DIQMTQSPSSLSASVGDRVTITCRASQ DVNTA VAWYQ QKP GKAPKLLIYSASFLYSGVPSRFSGSRSGTDFTLTISSLQPED FATYYCQQHYTTEPTF GQGTK VEIKRTVA A PS VFIFPPSDE QLKS GTASVVCLLNNFYPREAKVQWK VDNALQSGNSQES TEQDS KDSTYSLSSTLTLSKADYEKH KVYACEVTHQGLS 2 SPVTKSFNRGEC Proteinsekvens Anti-HER2 Heavy chain (1 och 2) PGKGLEWVARIYPTNGYTRYADS VKGRFTISADTSKNTAY LQM N S L R A E D T A V Y Y C S R w G G D G F Y A M D YwGQGTL VTV SSASTKGPSVEPLAPSSKSTSCGTAALGCLVKDYFPEPVTV SWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQT YICNVNHKPSNTKVDKK VEPKSCDKTHTCPPCPAPELLGOG PSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNW Y VDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQD WLNG KEYKCKVSNKALPAPIEKTISKAKGOPR EPQVYLPPSRE EMI T K N Q V S L T CLVKGFYPSDIAVEWESN GQPENNYKTTPP VLDSDGSFFLYSKLTVDKSRWQQGNVESCSVMHEALHNE YTQKSLSLSPGK

Explanation / Answer

Human epidermal growth factor receptor 2 which is also called HER2 or neu is an oncogene which involved in abnormal cell growth particular in breast cancer which is a target for the humanised monoclonal antibody trastuzumab (Herceptin).HER2/neu is generally overexpressed in 20–30% of patients who has metastatic breast cancer where due to this gene overexpression,disruption of normal signaling pathways occur which causing the loss of cell growth regulation and due to this development of resistance to apoptosis occur. Trastuzumab induces antibody-dependent cellular cytotoxicity which inhibits HER2 mediated signaling and prevents damage of the extracellular domain of HER2. In HER2-positive breast cancer drug trastuzumab has shown a survival advantage. Trastuzumab is produced by recombinant DNA technology in a mammalian cell mainly in Chinese Hamster Ovary culture.In laboratory,we examined whether transtuzumzb attributes are retained by the anti-HER2 Fc variants. WT and mutant anti-HER inhibited the growth of the HER2-overexpressing BT-474 and SK-BR-3 breast cancer cells,which is in a dose-dependent manner and we can compare potency to those of the clinically available trastuzumab (Herceptin). HEK293 is human embryonic kidney,which used for production of monoclonal antibody.In laboratory monoclonal antibodies produced by employing the cloning approach with the human embryonic kidney derivative cell line Expi293F. Antibody production with HEK293-F cells initiated by using 30mL shaker flask cultures under selection for 2weeks, which is followed by upto 2-week scale, up to 500mL shaker flask cultures. After 4weeks we obtained an average of 13µg/mL of anti-CSPG4 WT calculated by ELISA method.

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