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P2 biogel

Manufactured by Bio-Rad
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P2-BioGel is a size exclusion chromatography resin designed for the separation and purification of biomolecules. It is composed of agarose beads that provide a porous structure for the selective separation of molecules based on their size and shape.

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Lab products found in correlation

Dc protein assay kit 2, by Bio-Rad (2 mentions) D glucose, by Merck Group (1 mentions) Kta chromatography system, by GE Healthcare (1 mentions)

Spelling variants of this product

Biogel P2 Extrafine column (2 citations) BioGel P2 size exclusion column (2 citations) Size exclusion Biogel (P2) (2 citations) Biogel P2 Extrafine (2 citations)

8 protocols using p2 biogel

1

Oligosaccharide Conjugation via p-Nitrophenyl Ester

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To a solution of oligosaccharide (2–5 μmol) in 0.5 mL anhydrous dimethylformamide (DMF) solution was added p-nitrophenyl ester linker (10–25 μmol) and stirred for 5 h at room temperature. The reaction was monitored by thin layer chromatography (TLC) (5:3:1:1, MeOH /EtOAc /H2O /AcOH). The disappearance of free amine indicated completion of the reaction. The reaction mixture was evaporated under reduced pressure without heating to remove DMF. Reaction mixture was dissolved in 500 μl H2O, insoluble excess of linker was separated by filtration and water-soluble fraction was purified by P2 BIO-GEL (Bio-Rad) column chromatography and gradually eluted with H2O.The solution was then lyophilized to afford desired respective half esters (I-IV) as a light yellow colored solid.
Shivatare S.S., Rachel Cheng T.J., Cheng Y.Y., Shivatare V.S., Tsai T.I., Chuang H.Y., Wu C.Y, & Wong C.H. (2021). Immunogenicity evaluation of N-glycans recognized by HIV broadly neutralizing antibodies.. ACS chemical biology, 16(10), 2016-2025.
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2

Enzymatic Generation of Defined GA-AGP Oligosaccharides

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GA-AGP derived oligosaccharides were generated by incubating 20 g of the glycan with 1 µM of the β1,3-galactosidase BT0265 in 20 mM sodium phosphate buffer pH 7.0 implemented with 150 mM NaCl at 37 °C for 16 h. The oligosaccharide mixture was freeze dried and resuspended in water before being applied to a P2-BioGel (BioRad) column with a 0.22 ml/min flow rate. Fractions were evaluated for oligosaccharide content and purity by TLC. Pure fractions of defined oligosaccharides were pooled and concentrated. Oligosaccharide size was confirmed by Mass Spectrometry and HPAEC.
Cartmell A., Muñoz-Muñoz J., Briggs J., Ndeh D.A., Lowe E.C., Baslé A., Terrapon N., Stott K., Heunis T., Gray J., Yu L., Dupree P., Fernandes P.Z., Shah S., Williams S.J., Labourel A., Trost M., Henrissat B, & Gilbert H.J. (2018). Engineering a surface endogalactanase into Bacteroides thetaiotaomicron confers keystone status for arabinogalactan degradation. Nature microbiology, 3(11), 1314-1326.
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3

Bioactive Compound Purification and Screening

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Example 2

Five millilitres of CFSM were directly deposited on a P2 Biogel (Bio-Rad, Missasauga, ON., Canada) column (exclusion, 100 to 1,800 Da; 2.5×100 cm; Bio-Rad Laboratories Ltd.) and run at room temperature in 18-Ω water at a gravity flow rate of 0.8 ml/min, and eighty 5 ml fractions were collected. The fractions collected were freeze-dried and resuspended in 1 ml 18-Ω water for preliminary screening against EHEC LEE1, LEE2 and AI-2 production as previously described (29). The total protein content of the fractions was quantified using the BioRad DC protein assay kit II. Fractions showing a strong inhibitory activity against LEE expression and AI-2 production were selected.

US10716817B2. Antiviral methods and compositions comprising probiotic bacterial molecules (2020-07-21). University of Guelph [CA]. Inventors: Mansel Griffiths [CA].
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4

Galactooligosaccharide Isolation and Characterization

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Galactooligosaccharides were generated by incubation of 3 g of galactan with 100 mM HCl incubated for 3 h at 100 °C and neutralised by NaOH titration. The oligosaccharide mixture was freeze dried and resuspended in water before being applied to a P2-BioGel (BioRad) column with a 0.22 ml/min flow rate. Fractions were evaluated for oligosaccharide content and purity by TLC. Pure fractions of defined oligosaccharides were pooled and concentrated. Oligosaccharide size was confirmed by Mass Spectrometry and HPAEC. Crude oligosaccharide mixtures were generated by partial digestion with appropriate enzymes; BT0360 and BT0367 (arabinan), BT4668 (galactan), BT4170 (P-RGI/AM-RGI) and BT4116 (HG). Reactions were boiled and filter sterilised to remove precipitate before being evaluated by TLC.
Luis A.S., Briggs J., Zhang X., Farnell B., Ndeh D., Labourel A., Baslé A., Cartmell A., Terrapon N., Stott K., Lowe E.C., McLean R., Shearer K., Schückel J., Venditto I., Ralet M.C., Henrissat B., Martens E.C., Mosimann S.C., Abbott D.W, & Gilbert H.J. (2017). Dietary pectic glycans are degraded by coordinated enzyme pathways in human colonic Bacteroides. Nature microbiology, 3(2), 210-219.
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5

Enzymatic Generation of Defined GA-AGP Oligosaccharides

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GA-AGP derived oligosaccharides were generated by incubating 20 g of the glycan with 1 µM of the β1,3-galactosidase BT0265 in 20 mM sodium phosphate buffer pH 7.0 implemented with 150 mM NaCl at 37 °C for 16 h. The oligosaccharide mixture was freeze dried and resuspended in water before being applied to a P2-BioGel (BioRad) column with a 0.22 ml/min flow rate. Fractions were evaluated for oligosaccharide content and purity by TLC. Pure fractions of defined oligosaccharides were pooled and concentrated. Oligosaccharide size was confirmed by Mass Spectrometry and HPAEC.
Cartmell A., Muñoz-Muñoz J., Briggs J., Ndeh D.A., Lowe E.C., Baslé A., Terrapon N., Stott K., Heunis T., Gray J., Yu L., Dupree P., Fernandes P.Z., Shah S., Williams S.J., Labourel A., Trost M., Henrissat B, & Gilbert H.J. (2018). Engineering a surface endogalactanase into Bacteroides thetaiotaomicron confers keystone status for arabinogalactan degradation. Nature microbiology, 3(11), 1314-1326.
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6

CFSM Fractionation and Screening

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Example 2

Five millilitres of CFSM were directly deposited on a P2 Biogel (Bio-Rad, Missasauga, ON., Canada) column (exclusion, 100 to 1,800 Da; 2.5×100 cm; Bio-Rad Laboratories Ltd.) and run at room temperature in 18-Ω water at a gravity flow rate of 0.8 ml/min, and eighty 5 ml fractions were collected. The fractions collected were freeze-dried and resuspended in 1 ml 18-0 water for preliminary screening against EHEC LEE1, LEE2 and AI-2 production as previously described (29). The total protein content of the fractions was quantified using the BioRad DC protein assay kit II. Fractions showing a strong inhibitory activity against LEE expression and AI-2 production were selected.

US11857581B2. Antiviral methods and compositions comprising probiotic bacterial molecules (2024-01-02). MicroSintesis Inc. [CA]. Inventors: Mansel Griffiths [CA].
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7

Galactooligosaccharide Isolation and Characterization

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Galactooligosaccharides were generated by incubation of 3 g of galactan with 100 mM HCl incubated for 3 h at 100 °C and neutralised by NaOH titration. The oligosaccharide mixture was freeze dried and resuspended in water before being applied to a P2-BioGel (BioRad) column with a 0.22 ml/min flow rate. Fractions were evaluated for oligosaccharide content and purity by TLC. Pure fractions of defined oligosaccharides were pooled and concentrated. Oligosaccharide size was confirmed by Mass Spectrometry and HPAEC. Crude oligosaccharide mixtures were generated by partial digestion with appropriate enzymes; BT0360 and BT0367 (arabinan), BT4668 (galactan), BT4170 (P-RGI/AM-RGI) and BT4116 (HG). Reactions were boiled and filter sterilised to remove precipitate before being evaluated by TLC.
Luis A.S., Briggs J., Zhang X., Farnell B., Ndeh D., Labourel A., Baslé A., Cartmell A., Terrapon N., Stott K., Lowe E.C., McLean R., Shearer K., Schückel J., Venditto I., Ralet M.C., Henrissat B., Martens E.C., Mosimann S.C., Abbott D.W, & Gilbert H.J. (2017). Dietary pectic glycans are degraded by coordinated enzyme pathways in human colonic Bacteroides. Nature microbiology, 3(2), 210-219.
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8

SEC Separation of Saccharides

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The ÄKTA chromatography system (GE Healthcare, Chicago, United States) was used for the separation of the saccharides by SEC. A P-2 Bio-Gel (Bio-Rad Laboratories, Hercules, United States) column (5 × 175 mm) was manually prepared according to the manufacturer’s instruction. The flow rate was set to 0.1 ml/min, and deionized and degassed water was used for elution. Commercially available D-glucose and D-lactose monohydrate (both Merck KGaA Darmstadt, Germany), as well as the pentose verbascose (O-VER) (Megazyme Inc., Bray, Ireland), were used as standards. The saccharides were prepared as 20 mg/ml solutions in deionized water, and 200 μl was used for SEC separation. The lactase F-digested HMOs were resuspended in 100 μl deionized water and separated. Fractions of 200 μl were collected.
Jarzynka S., Spott R., Tchatchiashvili T., Ueberschaar N., Martinet M.G., Strom K., Kryczka T., Wesołowska A., Pletz M.W., Olędzka G, & Makarewicz O. (2022). Human Milk Oligosaccharides Exhibit Biofilm Eradication Activity Against Matured Biofilms Formed by Different Pathogen Species. Frontiers in Microbiology, 12, 794441.
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