Journal: PLoS ONE

Article Title: Robotic selection for the rapid development of stable CHO cell lines for HIV vaccine production

doi: 10.1371/journal.pone.0197656

Figure Lengend Snippet: (A) G418 selected colonies visible in a single 35mm well illuminated with white light at 6 days. (B) The same single 35mm well illuminated with 490 nm wavelength light. Colonies actively secreting rgp120 have a green “halo” visible at 525 nm. (C) Relative mean exterior fluorescence of halo for more than 10,000 colonies imaged by the ClonePix2 plotted by rank. The top ranking 0.1% of colonies (44) were robotically picked and cultured. The six clones expressing 0.2–0.4 g/L at day 56 are shown in red.

Article Snippet: To address these problems we combined a high efficiency electroporation device (MaxCyte STX, MaxCyte Inc., Gaithersburg, MD), a robotic cell selection system, ClonePix2 (Molecular Devices, Sunnyvale, CA) and a novel cell line (MGAT1 - CHO 3.4F10) that was recently developed in our lab [ ].

Techniques: Fluorescence, Cell Culture, Clone Assay, Expressing

Journal: PLoS ONE

Article Title: Robotic selection for the rapid development of stable CHO cell lines for HIV vaccine production

doi: 10.1371/journal.pone.0197656

Figure Lengend Snippet: 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 μ l 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).

Article Snippet: To address these problems we combined a high efficiency electroporation device (MaxCyte STX, MaxCyte Inc., Gaithersburg, MD), a robotic cell selection system, ClonePix2 (Molecular Devices, Sunnyvale, CA) and a novel cell line (MGAT1 - CHO 3.4F10) that was recently developed in our lab [ ].

Techniques: Western Blot, Affinity Purification, Produced, Purification, Enzyme-linked Immunosorbent Assay, SDS Page, SDS-Gel, Staining