科研之友
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摘要
The baculovirus vector expression system is considered a useful insect cell-based platform for large-scale production of recombinant biopharmaceutical glycoproteins. The GA733 antigenic protein is a 40 kDa cell surface glycoprotein that is overexpressed in human colorectal carcinoma. Three GA733-expressing baculovirus vectors were constructed: (i) GA733 fused to the Fc fragment of human immunoglobulin G (GA733-Fc); (ii) GA733-Fc tagged with the endoplasmic reticulum (ER) retention signal KDEL (GA733-FcK); and (iii) GA733-FcK without the signal peptide [GA733-FcK (w/o SP)]. These three recombinant proteins were expressed in Fall Armyworm (Spodoptera frugiperda) Sf9 insect cells. GA733-Fc purified from insect culture medium (CM) and cell lysate (CL) (GA733(I)-Fc(CM) and GA733(I)-Fc(CL), respectively), and GA733-FcK and GA733-FcK (w/o SP) purified from CL [GA733(I)-Fc(CL) and GA733(I)-FcK(CL) (w/o SP), respectively] were injected intraperitoneally into BALB/c mice [three times at 1 mu g, with or without an adjuvant (aluminum hydroxide)]. Surface plasmon resonance (SPR) analysis (involving a chip with the immobilized GA733 antigen) showed that, in general, the serum samples from mice immunized with the adjuvant yielded stronger SPR signals than did those from mice immunized without an adjuvant. The serum samples from mice after the second injection yielded stronger signals than did those from mice before the second injection. The group immunized with GA733(I)-FcK(CL) (glycosylated with an oligomannose) showed the strongest SPR signals. The SPR signals from the GA733(I)-FcK(CL) group were stronger than or similar to those from mice immunized with mammalian-cell-expressed GA733-Fc (GA733(M)-Fc). Thus, the baculovirus vector expression system can be used to produce immunogenic cancer glycoprotein.
- 出版日期2016-1
