mode resins cht type ii  (Bio-Rad)

Journal: Protein expression and purification

Article Title: A scalable and high yielding SARS-CoV-2 spike protein receptor binding domain production process.

doi: 10.1016/j.pep.2023.106241

Figure Lengend Snippet: Fig. 5. Milligram Scale CHT Chromatography and Corresponding HPSEC Profile for SARS-CoV-2 Spike Protein RBD. 5A shows the chromatography profile of SARS- CoV-2 Spike Protein RBD from the CHT resin. Flow through, wash, and strip peaks were observed. 5B, 5C, 5D, and 5E respectively show the flow through, wash 1, wash 2, and strip HPSEC purity profiles. UV A280, conductivity, and pH traces are respectively shown in blue, orange, and purple.

Article Snippet: Development of a scalable aggregate removal operation to replace the SEC operation consisted of studying the behavior of SARS-CoV-2 Spike Protein RBD from the Wuhan strain on CHT Type II resin (BIORAD; Hercules, CA, USA) resin (1.6 cm column diameter, 14.8 cm bed height).

Techniques: Chromatography, Stripping Membranes

Journal: Protein expression and purification

Article Title: A scalable and high yielding SARS-CoV-2 spike protein receptor binding domain production process.

doi: 10.1016/j.pep.2023.106241

Figure Lengend Snippet: Fig. 8. SDS-PAGE and Western blot Analyses of Gram Scale SARS-CoV-2 Spike Protein RBD. The CHT Type II gram scale product was analyzed by SDS-PAGE in 8A and Western blot in 8B. The milligram control and CHT Flow Through Products were visualized by reaction with a biotinylated polyclonal antibody specific for the poly-histidine tag prior to application of a streptavidin labeled peroxidase. Extraneous lanes were removed from the image.

Article Snippet: Development of a scalable aggregate removal operation to replace the SEC operation consisted of studying the behavior of SARS-CoV-2 Spike Protein RBD from the Wuhan strain on CHT Type II resin (BIORAD; Hercules, CA, USA) resin (1.6 cm column diameter, 14.8 cm bed height).

Techniques: SDS Page, Western Blot, Control, Labeling

Journal: Protein expression and purification

Article Title: A scalable and high yielding SARS-CoV-2 spike protein receptor binding domain production process.

doi: 10.1016/j.pep.2023.106241

Figure Lengend Snippet: Fig. 7. Gram Scale CHT Elution and Corresponding HPSEC Profile for SARS-CoV-2 Spike Protein RBD. 7A shows the elution profile of SARS-CoV-2 Spike Protein RBD from CHT Type II resin. 7B, 7C, and 7D contain the respective HPSEC purity profiles of the flow through, re-equilibration (Wash 1), and high phosphate (Wash 2) CHT products. The UV A280, conductivity, and pH traces are respectively shown in blue, orange, and purple.

Article Snippet: Development of a scalable aggregate removal operation to replace the SEC operation consisted of studying the behavior of SARS-CoV-2 Spike Protein RBD from the Wuhan strain on CHT Type II resin (BIORAD; Hercules, CA, USA) resin (1.6 cm column diameter, 14.8 cm bed height).

Techniques:

Journal: Protein expression and purification

Article Title: A scalable and high yielding SARS-CoV-2 spike protein receptor binding domain production process.

doi: 10.1016/j.pep.2023.106241

Figure Lengend Snippet: Fig. 11. CHT Elution and Corresponding Purity Profiles of SARS-CoV-2 Omicron BA.2 Spike Protein RBD. 11A shows the elution profile of SARS-CoV-2 Omicron Spike Protein RBD from CHT Type II resin. 11B, 11C, 11D, and 11E contain the respective HPSEC purity profiles of the flow through, re-equilibration (Wash 1), and high salt (Strip1) and high phosphate (Strip 2) CHT products. The UV A280, conductivity, and pH traces are respectively shown in blue, orange, and purple. 11F shows the SDS-PAGE analysis of the monomeric flow through product.

Article Snippet: Development of a scalable aggregate removal operation to replace the SEC operation consisted of studying the behavior of SARS-CoV-2 Spike Protein RBD from the Wuhan strain on CHT Type II resin (BIORAD; Hercules, CA, USA) resin (1.6 cm column diameter, 14.8 cm bed height).

Techniques: Stripping Membranes, SDS Page

Journal: bioRxiv

Article Title: ATP-binding cassette protein ABCF1 couples gene transcription with maintenance of genome integrity in embryonic stem cells

doi: 10.1101/2020.05.28.122184

Figure Lengend Snippet: (A) Chromatography scheme for purification of SCC-B from NT2 nuclear extracts (NT2 NE). NT2 NE is first subjected to ammonium sulfate precipitation (55% saturation) followed by a series of chromatographic columns including nickel affinity agarose (Ni-NTA), cation exchangers phosphocellulose (P11), heparin (Poros-HE), Mono S, anion exchanger Poros-HQ, hydroxyapatite (HAP), and gel filtration medium Superose 6. SCC-B activity segregates from SCC-A (Dyskerin (DKC1) RNP) at the Poros-HE step. (B) Input fraction containing SCC-B activity from the Poros-HE step (IN), flow-through (FT) and various salt-eluted Mono S fractions were assayed for their ability to stimulate OCT4/SOX2-dependent transcription from the human NANOG promoter template engineered with four extra copies of the oct-sox composite binding element (bottom). All reactions contain purified general transcription factors (GTFs), Pol II, OCT4, SOX2, and recombinant XPC and DKC1 complexes. (C) Fractions assayed in vitro transcription shown in (B) are separated on a 4-12% gradient polyacrylamide gel and stained with silver. Filled arrowhead indicates the predominant polypeptide at ~110 kDa that co-migrates with SCC-B transcriptional activity. The following figure supplement is available for : .

Article Snippet: Transcriptionally active Q0.3 fraction (0.32–0.4 M) were pooled and applied directly to hydroxyapatite (HAP) type II ceramic resin (Bio-Rad), washed first at 0.38 M, then lowered to 0.1 M KCl in 3 CV.

Techniques: Chromatography, Purification, Filtration, Activity Assay, Binding Assay, Recombinant, In Vitro, Staining