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Antibody purification resins—an alternative to Protein A

Antibody purification resins

Antibody purification using thiophilic resins is a simple, powerful, and economical alternative to Protein A, the most widely used method of immunoglobulin purification. Although Protein A antibody purification is very common, there are certain types of antibodies, such as the single-chain antibodies IgE, IgY, and IgM, that cannot be purified using Protein A. An alternative antibody purification method, thiophilic affinity chromatography (TAC), is ideal for these types of applications, as well as immunoglobulin purification in general, including IgG, IgM, IgA, Fab and Fc fragments, and C3 and C4 complement factors.

Antibody purification using thiophilic resins is a simple, powerful, and economical alternative to Protein A, the most widely used method of immunoglobulin purification. Although Protein A antibody purification is very common, there are certain types of antibodies, such as the single-chain antibodies IgE, IgY, and IgM, that cannot be purified using Protein A. An alternative antibody purification method, thiophilic affinity chromatography (TAC), is ideal for these types of applications, as well as immunoglobulin purification in general, including IgG, IgM, IgA, Fab and Fc fragments, and C3 and C4 complement factors.

TAC is a powerful and simple method for the purification of proteins—in particular, immunoglobulins. It is an economical alternative to Protein A for purifying antibodies from whole serum and tissue culture. In comparison to Protein A-based immunoadsorbents, thiophilic adsorbents have a broader affinity for immunoglobulins. Further, more than 99% of total proteins are recovered using a thiophilic adsorbent, in comparison to less than 92% for phenyl and 75% for octyl agarose adsorbents.

TAC has broad specificity for immunoglobulins of different classes and subclasses from a wide range of species and has capacity similar to that of other affinity methods.

Its advantages are the simplicity of the sample preparation, applicability with a range of source materials and protein concentration ranges, and gentle elution conditions, unlike the popular Protein A/Protein G separation methods that require extreme pH for elution, which the antibodies may not be stable enough to accommodate.

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Cat. # Product Size Price License Quantity Details
635616 Thiophilic-Superflow Resin 10 mL USD $282.00

Thiophilic-Superflow Resin is an adsorbent for large-scale or FPLC purifcation of various proteins, including IgG, IgM, IgY, and IgE antibodies. Superflow allows for flow rates of as high as 5 ml/cm2 min.

Notice to purchaser

Our products are to be used for Research Use Only. They may not be used for any other purpose, including, but not limited to, use in humans, therapeutic or diagnostic use, or commercial use of any kind. Our products may not be transferred to third parties, resold, modified for resale, or used to manufacture commercial products or to provide a service to third parties without our prior written approval.

Documents Components Image Data

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635616: Thiophilic-Superflow Resin

635616: Thiophilic-Superflow Resin

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Structure of Thiophilic Resin

Structure of Thiophilic Resin
Structure of Thiophilic Resin.

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Purification using Thiophilic Resin vs Protein A

Purification using Thiophilic Resin vs Protein A
Purification using Thiophilic Resin vs Protein A. Purification with Thiophilic Resin allows elution in any buffer with pH 7.0. In contrast, Protein A requires elution in a buffer with pH lower than 7.0. Protein A also requires a dialysis step, which can be omitted with Thiophilic Resin.

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Single-chain antibody purification with Thiophilic Resin

Single-chain antibody purification with Thiophilic Resin
Single-chain antibody purification with Thiophilic Resin. SDS-PAGE analysis of the following samples: bacterial lysate expressing scFv215 (Lane 2), periplasmic fraction (Lane 3), peak fraction from Ni-NTA (Lane 4), and peak fraction from Thiophilic Resin (Lane 5). (Schulze, R. A. et al. (1994) Anal. Biochem. 220(1):212–214. Permission to reprint obtained.).

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IgY purification from chicken egg using Thiophilic Resin

IgY purification from chicken egg using Thiophilic Resin
IgY purification from chicken egg using Thiophilic Resin. Panel A. SDS-PAGE analysis of fractions from purification of chicken egg immunoglobulins. Lanes 7 & 10 each contain 10 mg of protein, and all other lanes contain 25 mg each. Lane 1: egg yolk extract supernatant. Lane 2: Supernatant after 60% SAS (Saturated concentration of Ammonium Sulfate). Lane 3: Wash with 60% SAS. Lane 4: Pellet after 60% SAS. Lane 5: column load. Lane 6: unbound material. Lane 7: eluted material. Lanes 8–10: purification with another thiophilic resin. Panel B. Immunoblot using 1/10 the amount of material in Panel A. IgY was detected with polyclonal rabbit anti-chicken HRP-conjugate. M=molecular weight. (Hansen, P. et al. (1998). J. Immun. Methods 215(1–2):1–7. Permission to reprint obtained).
635617 Thiophilic-Superflow Resin 100 mL USD $1763.00

Thiophilic-Superflow Resin is an adsorbent for large-scale or FPLC purifcation of various proteins, including IgG, IgM, IgY, and IgE antibodies. Superflow allows for flow rates of as high as 5 ml/cm2 min.

Notice to purchaser

Our products are to be used for Research Use Only. They may not be used for any other purpose, including, but not limited to, use in humans, therapeutic or diagnostic use, or commercial use of any kind. Our products may not be transferred to third parties, resold, modified for resale, or used to manufacture commercial products or to provide a service to third parties without our prior written approval.

Documents Components Image Data

Back

Structure of Thiophilic Resin

Structure of Thiophilic Resin
Structure of Thiophilic Resin.

Back

Purification using Thiophilic Resin vs Protein A

Purification using Thiophilic Resin vs Protein A
Purification using Thiophilic Resin vs Protein A. Purification with Thiophilic Resin allows elution in any buffer with pH 7.0. In contrast, Protein A requires elution in a buffer with pH lower than 7.0. Protein A also requires a dialysis step, which can be omitted with Thiophilic Resin.

Back

Single-chain antibody purification with Thiophilic Resin

Single-chain antibody purification with Thiophilic Resin
Single-chain antibody purification with Thiophilic Resin. SDS-PAGE analysis of the following samples: bacterial lysate expressing scFv215 (Lane 2), periplasmic fraction (Lane 3), peak fraction from Ni-NTA (Lane 4), and peak fraction from Thiophilic Resin (Lane 5). (Schulze, R. A. et al. (1994) Anal. Biochem. 220(1):212–214. Permission to reprint obtained.).

Back

IgY purification from chicken egg using Thiophilic Resin

IgY purification from chicken egg using Thiophilic Resin
IgY purification from chicken egg using Thiophilic Resin. Panel A. SDS-PAGE analysis of fractions from purification of chicken egg immunoglobulins. Lanes 7 & 10 each contain 10 mg of protein, and all other lanes contain 25 mg each. Lane 1: egg yolk extract supernatant. Lane 2: Supernatant after 60% SAS (Saturated concentration of Ammonium Sulfate). Lane 3: Wash with 60% SAS. Lane 4: Pellet after 60% SAS. Lane 5: column load. Lane 6: unbound material. Lane 7: eluted material. Lanes 8–10: purification with another thiophilic resin. Panel B. Immunoblot using 1/10 the amount of material in Panel A. IgY was detected with polyclonal rabbit anti-chicken HRP-conjugate. M=molecular weight. (Hansen, P. et al. (1998). J. Immun. Methods 215(1–2):1–7. Permission to reprint obtained).

Back

635617: Thiophilic-Superflow Resin

635617: Thiophilic-Superflow Resin

Overview

Thiophilic-Superflow Resin utilizes thiophilic adsorption chromatography. With this technique, protein adsorbs to a sulfone thioether ligand. The adsorption of different proteins can be promoted by adding different salts to the mixture. Varying the concentration of the loading salt can affect the adsorption affinities of IgG, IgM, IgA, Fab and Fc fragments, and C3 and C4 complement factors.

Purification with Thiophilic-Superflow Resin offers several advantages over purification with Protein A, the conventional method used for immunoglobulin purification:

  • Purification at neutral pH
  • Broader selectivity (IgY, IgM, IgE, scAb)
  • High linear flow rates
  • High capacity (15–20 mg Ab/ml resin)
  • Reusability
  • Higher stability of the purified product

More Information

Applications

  • Antibody purification, including IgG, IgM, and chicken IgY

Properties of Thiophilic-Superflow Resin

Feature Thiophilic-Superflow Resin
Batch/gravity Yes
FPLC Yes
Scale Analytical, preparative
Preparative production capacity (mg IgG/ml absorbent) 25
Matrix Cross-linked agarose
Maximum linear flow rate (cm/min) 5.0
Maximum volumetric flow rate (ml/min) at 5 x 1 cm i.d. 4.0
pH stability 2–10
Supplied as Bulk/slurry 50% in 25% ethanol
Storage 4°C, do not freeze


Additional product information

Please see the product's Certificate of Analysis for information about storage conditions, product components, and technical specifications. Please see the Kit Components List to determine kit components. Certificates of Analysis and Kit Components Lists are located under the Documents tab.


An efficient, versatile, and economical alternative to Protein A purification

Thiophilic antibody purification resins

An efficient, versatile, and economical alternative to Protein A purification.

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