america type culture collection tib 202 crl 2019  (ATCC)

Journal: bioRxiv

Article Title: A Prefusion Form of Herpes Simplex Virus 1 gB has a Distinct Antigenic Signature

doi: 10.64898/2026.04.12.718011

Figure Lengend Snippet: (A) CHO-K1 effector cells transiently expressing wild type gB or gB H516P and wild type gD, gH, and gL, and T7 polymerase were co-cultured for 18 h with CHO-HVEM cells transiently expressing the luciferase gene. Luciferase-induced luminescence was quantitated as a measure of fusion. (B) Detection of gB on the cell surface by CELISA. CHO-K1 cells were transfected with plasmids encoding HSV-1 gB or empty vector for 24 h. Cells were fixed with paraformaldehyde and then incubated with anti-gB monoclonal antibodies H126, H1359, and H1817. HRP-conjugated Protein A was added, followed by ABTS substrate. Absorbance was read at 405 nm. Data was normalized to wild type gB set to 100% for both experiments. Results are the means of three independent experiments. *, p < 0.05; ns, not significant, Student’s t- test.

Article Snippet: Chinese hamster ovary (CHO-K1) cells (American Type Culture Collection (ATCC), Manassas, VA, USA) were propagated in Ham’s F12 nutrient mixture (Gibco/Life Technologies, Grand Island, NY, USA) supplemented with 10% fetal bovine serum (FBS) (Atlanta Biologicals, Atlanta, GA, USA).

Techniques: Expressing, Cell Culture, Luciferase, Transfection, Plasmid Preparation, Incubation, Bioprocessing

Journal: bioRxiv

Article Title: A Prefusion Form of Herpes Simplex Virus 1 gB has a Distinct Antigenic Signature

doi: 10.64898/2026.04.12.718011

Figure Lengend Snippet: (A) Space-filling rendering (PyMOL) of prefusion gB (PDB 6Z9M) and postfusion gB (PDB 2GUM) ectodomain trimer . Epitopes of monoclonal antibodies used in this study are highlighted in color. The MAb H126 epitope includes residue 303 in gB domain I (blue) ( , ). The MAb H1838 epitope maps to residues 391 – 410 in domain II (green) . The H1359 epitope maps to residue 487-505 in domain III (yellow) . The SS10 epitope maps to residues 640 – 670 in domain IV (orange) . The SS106 epitope maps to residue 697 – 725 in domain V (red). The MAb DL16 epitope is trimer-specific and resides in domain V . The MAb H1817 epitope maps to residues 31 – 43 in gB domain VI (white) , which is unresolved in the structure. (B-H) gB H516P is antigenically distinct from wild type gB at domains II, IV, and V. Lysates of CHO-K1 cells expressing wild type gB (red) or H516P gB (purple) were bound to nitrocellulose membrane then probed with antibodies to gB. The appropriate fluorescent-conjugated secondary antibody was added for 20 min. Images were generated with Azure Biosystems imager. Rabbit polyclonal antibody R68 was used to standard loading. Reactivity of the antibodies was quantitated relative to the wild type by densitometry using Image Studio. The values shown represent antibody reactivity relative to wild type gB set to 100%. Results are the means and standard deviations of three independent experiments. ** p < 0.005; *** p < 0.001, Student’s t- test. (I) Summary of the antigenic analysis of gB H516P. The slopes of the lines of best fit for H516P gB relative to wild type (panels B-H) were calculated and used to determine fold changes, which were normalized to 1.0.

Article Snippet: Chinese hamster ovary (CHO-K1) cells (American Type Culture Collection (ATCC), Manassas, VA, USA) were propagated in Ham’s F12 nutrient mixture (Gibco/Life Technologies, Grand Island, NY, USA) supplemented with 10% fetal bovine serum (FBS) (Atlanta Biologicals, Atlanta, GA, USA).

Techniques: Bioprocessing, Residue, Expressing, Membrane, Generated

Journal: bioRxiv

Article Title: A Prefusion Form of Herpes Simplex Virus 1 gB has a Distinct Antigenic Signature

doi: 10.64898/2026.04.12.718011

Figure Lengend Snippet: Lysate from CHO-K1 cells expressing wild type gB or H516P gB was treated at a range of pH values (7.4 to 5.0) for 10 min at 37°C. Samples were blotted to nitrocellulose membranes and probed at neutral pH with gB MAbs targeting epitopes undergoing conformational change in (A) Domain 1, H126; (B) Domain V, SS106; and epitope not undergoing conformational change in (C) Domain IV, SS10 at neutral pH. Fluorescent-conjugated secondary antibody was added for 20 min. Images were generated with Azure Biosystems imager. Reactivity of three independent experiments was quantified by densitometry. The pH 7.4-treated samples were set to 100%.

Article Snippet: Chinese hamster ovary (CHO-K1) cells (American Type Culture Collection (ATCC), Manassas, VA, USA) were propagated in Ham’s F12 nutrient mixture (Gibco/Life Technologies, Grand Island, NY, USA) supplemented with 10% fetal bovine serum (FBS) (Atlanta Biologicals, Atlanta, GA, USA).

Techniques: Expressing, Generated

Journal: bioRxiv

Article Title: A Prefusion Form of Herpes Simplex Virus 1 gB has a Distinct Antigenic Signature

doi: 10.64898/2026.04.12.718011

Figure Lengend Snippet: (A) CHO-K1 cells were transfected with plasmids encoding wild type gB or H516P gB. Cell lysates were either prepared in 2% SDS sample buffer with reducing agent and heated (denatured conditions), and then resolved by SDS-PAGE (left panel) or prepared with 0.2% SDS, no reducing agent, no heating (“native” conditions) and resolved by SDS-PAGE (right panel). Western blots were probed with anti-gB polyclonal antibody R68. Molecular weight standards (kDa) are indicated to the right. β-actin was included as a loading control. (B) Lysates of CHO-K1 cells expressing wild type gB (red) or H516P gB (purple) were bound to nitrocellulose membrane then probed with MAb DL16. Anti-mouse fluorescent-conjugated secondary antibody was added for 20 min. Images were generated with Azure Biosystems imager. Reactivity of DL16 was quantitated relative to the wild type by densitometry using Image Studio. gB wild type, 1.00; gB H516P, 0.31+/-0.01****. The values shown represent antibody reactivity relative to wild type gB set to 100%. Results are the means and standard deviations of three independent experiments. *** p < 0.0001, Student’s t- test.

Article Snippet: Chinese hamster ovary (CHO-K1) cells (American Type Culture Collection (ATCC), Manassas, VA, USA) were propagated in Ham’s F12 nutrient mixture (Gibco/Life Technologies, Grand Island, NY, USA) supplemented with 10% fetal bovine serum (FBS) (Atlanta Biologicals, Atlanta, GA, USA).

Techniques: Transfection, SDS Page, Western Blot, Molecular Weight, Control, Expressing, Membrane, Generated