Robotic selection for the rapid development of stable CHO cell lines for HIV vaccine production
Fig 3
Analysis of A244_N332-rgp120 secreted from stable MGAT1- CHO cell lines.
Six stable MGAT1- CHO cell lines identified with the ClonePix2 were selected as potential substrates for HIV vaccine production. (A) Immunoblot of affinity-purified rgp120 (50 ng per lane) produced by each of six A244_N332-rgp120 cell lines: 3E, 5C, 5D, 3F, 6B, and 5F. Purified A244_N332-rgp120 produced in normal CHO DG44 cells (692) was shown for purpose of comparison. (B) Comparison of A244_N332-rgp120 protein yields as determined by ELISA from the six MGAT1- CHO cell lines. (C) SDS PAGE of rgp120 produced by the 5F MGAT1- CHO cell line. Supernatant samples (10 μl per lane) collected over the time course of the culture were electrophoresed on a 4–12% NuPage PAGE SDS gel in MOPS buffer (Thermo Scientific, Waltham, MA). The gel was stained with Simply Blue (Thermo Scientific, Waltham, MA) and visualized using an Innotech FluoChem2 system (Genetic Technologies, Grover, MO).