The establishment of the production process for recombinant proteins expressed in mammalian cells : dissertation thesis = Postavitev procesa za produkcijo rekombinantnih beljakovin eksprimiranih v celicah sesalcev : doktorska disertacija
Knežević, Miomir
Gen za humani interleukin-2 (IL-2), ki je baziral na cDNA smo klonirali v ekspresijskivektor pRc/RSV. Stem konstruktom (pRc/RSV SIGIL2) smo transficirali hrčkove ledvične celice (BHK-21) in ovarijske ...celice hrčka (CHOdhfr-). Specifična produktivnost IL2 za klon BIL2T2H11 se je gibala od 4-10 ng/10 g celic/dan. Rekombinantni interleukin-2 je v odvisnosti od ekspresijskega sistema vplival na kinetiko rasti CTLL-2 celic. Bakterijski produkt je pri povišani koncentraciji deloval zaviralno, medtem ko je rBHK IL2 tudi pri višjih koncentracijah deloval ugodno na rast CTLL-2 celic. Razlika med rekombinantnima interleukinoma se je pokazala v začetnem in končnem delu gojitve CTLL-2 celic, saj bakterijski IL2 deluje hitreje, toda ob koncu poskusa rBHK IL2 bolj stimulira rast testnih celic. Izbor najugodnejšega nosilca BIL2T2H11 celično linijo je bil narejen na osnovi študije sposobnosti imobilizacije BHK starševske linije na nosilcih: Cytodex 3 (Pharmacia), Siran (Shott), Microsphere (Verax), PS80400, PE1 in PE2. Mikroporozni celični nosilec PE2 se je izkazal kot najprimernejši tako v mediju z 2-5% FCS (fetalnem telečjem serumu) kakor tudi v brezproteinskem mediju.
Type of material - dissertation
Publication and manufacture - Ljubljana : [M. Knežević], 1995
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