Protein L Resin

Produktnummer ABIN1536561

Produktdetails

Target: Protein L

Reaktivität: Peptostreptococcus magnus

Applikation: Affinity Chromatography (AC), Purification (Purif)

Verwendungszweck: Protein L Resin is an affinity chromatography medium designed for easy, one-step purification of classes, subclasses and fragments of immunoglobulins from biological fluids and from cell culture media.

Spezifität: Ligand Highly purified protein L
Number of Ig binding sites per ligand 5
MW of ligand Approximately 42 kDa
PI of ligand 4.57
Degree of substitution Approximately 2 mg protein L/ml settled resin
Static binding capacity >15 mg rabbit IgG/ml settled resin
Matrix spherical Agarose, 4% cross-linked
Average particle size 90 μm (45 - 165 μm)
Storage solution 1×PBS containing 20% ethanol
Sanitization Washing of the packed column with 70% ethanol

Produktmerkmale: The highly purified protein L ligand is coupled to 4% highly cross-linked agarose. The coupling is optimized to give high binding capacity for immunoglobulins. The static binding capacity of Protein L Resin is greater than 15 mg rabbit IgG/ml settled resin. The dynamic binding capacity will vary depending on several factors such as target antibody, flow rate etc.

Bead Ligand: Protein L

Bead Matrix: Agarose beads

Bead Size: 90 µm

Anwendungsinformationen

Kommentare:

High temperature heating is not recommended. The agarose melts above 65°C.

Aufbereitung der Reagenzien:

Water and chemicals used for buffer preparation should be of high purity. It is recommended filtering the buffers by passing them through a 0.45 μm filter before use.
Binding/Wash Buffer: 20 mM Na2HPO4, 0.15 M NaCl, pH 8.0
Elution Buffer: 0.1 M glycine, pH 2.5
Neutralization Buffer: 1 M Tris-HCl, pH 8.5

Aufbereitung der Proben:

To insure that proper ionic strength and pH are maintained for optimal binding, it is necessary to dilute serum samples, ascite fluid or cell culture supernatant at least 1:1 with Binding/Wash Buffer. Alternatively, the sample may be dialyzed overnight against Binding/Wash Buffer.

Testdurchführung:

Packing of Column
1) Resuspend completely the resin and transfer 1 ml slurry to a new column, in which 1 ml Binding/Wash Buffer was added in advance.
2) Allow the resin to settle down and the buffer to drain from the column.
3) Add 5 ml binding/Wash Buffer onto the column to equilibrate the resin and drain the buffer with a flow speed of about 1 ml/min.

Column Purification
1) Apply the sample onto the column and drain the flow-through with a flow speed of about 1 ml/min. Collect the flow-through for measuring the binding efficiency to the resin, i.e. by SDS-PAGE.
2) Wash the column with 30 ml Binding/Wash Buffer and drain the buffer with a flow speed of about 2 ml/min, or until the absorbance of the effluent at 280 nm is stable.
3) Elute the immunoglobulins with 10-15 ml Elution Buffer and drain the eluate with a flow speed of about 1 ml/min. Collect the eluate and immediately neutralize to pH 7.4 with Neutralization Buffer (1/10 volume of total eluate)

Regeneration of Column
Regenerate the column by washing the resin with 10 ml Elution Buffer followed by equilibration with 5 ml Binding/Wash Buffer. Columns can be regenerated up to 10 times without significant loss of binding capacity.

Beschränkungen: Nur für Forschungszwecke einsetzbar

Handhabung

Format: Liquid

Lagerung: 4 °C

Informationen zur Lagerung: Store regenerated Protein L Resin in Binding/Wash Buffer containing 20% ethanol at 2°C to 8°C. Do not freeze.

Haltbarkeit: 18 months

Referenzen

Safdari, Farajnia, Asgharzadeh, Omidfar, Khalili: "humMR1, a highly specific humanized single chain antibody for targeting EGFRvIII." in: International immunopharmacology, Vol. 18, Issue 2, pp. 304-10, (2014) (PubMed).

De Oliveira, Wang, Ryan, Morrison, Kohn, Hollis: "A CD19/Fc fusion protein for detection of anti-CD19 chimeric antigen receptors." in: Journal of translational medicine, Vol. 11, pp. 23, (2013) (PubMed).

Zhang, Mao, Cao, Xiong, Wen, Chen, Zhu: "Generation and characterization of a novel recombinant antibody against LMP1-TES1 of Epstein-Barr virus isolated by phage display." in: Viruses, Vol. 5, Issue 4, pp. 1131-42, (2013) (PubMed).

Lin, Zhang, Wang, Lu, Zheng, Wang, Tang, Xu, Chen, Zhang, Zhao, Zhu, Mao, Feng: "A novel human Fab antibody for Trop2 inhibits breast cancer growth in vitro and in vivo." in: International journal of cancer. Journal international du cancer, Vol. 134, Issue 5, pp. 1239-49, (2013) (PubMed).

Chen, Zhang, Mao, Zhu, Ming, Wen, Ma, Cao, Lin, Tang, Liang, Feng: "A human Fab-based immunoconjugate specific for the LMP1 extracellular domain inhibits nasopharyngeal carcinoma growth in vitro and in vivo." in: Molecular cancer therapeutics, Vol. 11, Issue 3, pp. 594-603, (2012) (PubMed).

Rouet, Lowe, Dudgeon, Roome, Schofield, Langley, Andrews, Whitfeld, Jermutus, Christ: "Expression of high-affinity human antibody fragments in bacteria." in: Nature protocols, Vol. 7, Issue 2, pp. 364-73, (2012) (PubMed).

Wang, Zhang, Liu, Chen, Feng, Shang, Maziarz, Radtke, Hampe: "Anti-idiotypic antibody specific to GAD65 autoantibody prevents type 1 diabetes in the NOD mouse." in: PLoS ONE, Vol. 7, Issue 2, pp. e32515, (2012) (PubMed).

Dudgeon, Rouet, Kokmeijer, Schofield, Stolp, Langley, Stock, Christ: "General strategy for the generation of human antibody variable domains with increased aggregation resistance." in: Proceedings of the National Academy of Sciences of the United States of America, Vol. 109, Issue 27, pp. 10879-84, (2012) (PubMed).

Antigendetails

Target: Protein L

Abstract: Protein L Produkte

Hintergrund: Protein L, first isolated from the surface of bacterial species Peptostreptoccus magnus, binds immunoglobulin through κ light chain interaction. Protein L binds a wider range of Ig classes and subclasses than other antibody-binding proteins such as protein A or protein G. It can bind to all classes of Ig (i.e., IgG, IgM, IgA, IgE, and IgD) and also the single chain variable fragments (Scfv) and Fab fragments.