Protein A and Protein G are widely used at present for the purification of IgG from various animal species. However, the use of Protein A and Protein G presents certain inconveniences:
- These proteins are expensive.
- Protein A and Protein G do generally not bind IgM.
- The elution at acid pH results in a partial denaturation of IgG.
- These proteins do not recognize IgG from all animal species such as dog, chicken...

AFFILAND has developed a Thiophilic gel (T-Gel) which allows the purification of immunoglobulins from all animal species under mild conditions (pH 8.0, low salt concentration, elution at physiological pH)^**.

CHARACTERISTICS OF THE THIOPHILIC GEL or T-GEL

• This matrix (Agarose 4% cross-linked) contains thioether groups. The interactions of the immobilized thioether groups on the surface of the gel result in the formation of "pseudo S-S bridges" analogous to those present on the F_c fragment of immunoglobulins. These S-S structure interactions could explain the specific binding of immunoglobulins to the matrix.
• The ligand does not show any interactions with serum proteins other than immunoglobulins (plus traces of alpha2-macroglobulin which are easily removed on PDC-Agarose). Of special interest is that no trace of serum albumin binding to the T-Gel can be detected.
• The immunoglobulin capacity of the T-Gel is between 10 and 22 mg/ml gel. This capacity was observed for all animal species studied to date.
• The coating of mouse monoclonal IgG purified by AFFILAND T-gel shows negligible non-specific binding in IRMA (immunoradiometric assay).
• The gel can be easily regenerated.
• The gel can be reused at least 10 times with no loss of binding capacity.
• Stability of the gel: at 20°C for minimum a week; pH 2-10.
• Shelf life of the gel: several years if stored at 4°C.
• Storage: 4°C in PBS 50 mM pH 7.4; NaN₃ 0.02% (w/v).

ORDERING INFORMATION

Product code	Product name	Package	*Price in EUR
T-56-4S	T-Gel on SepharoseTM CL-4B	25 ml of gel	245.00
		50 ml of gel	409.00
		100 ml of gel	682.00
		500 ml of gel	2455.00
T-56-4FF	T-Gel on SepharoseTM 4 Fast Flow	25 ml of gel	320.00
		50 ml of gel	532.00
		100 ml of gel	886.00
		500 ml of gel	3190.00

* All prices are FOB Liège.
Bulk quantities are available.

IMMUNOGLOBULIN PURIFICATION BY T-GEL

Instruction for use

Gel volume: 10 ml T-GEL.
Sample¹: 50 ml serum from all animal species + binding buffer.
Binding buffer: (NH₄)₂SO₄ 0.9M pH 8.0 (with NaOH 1M): buffer A.
Flow rate: 40ml/cm²/hour for all operations.
Washing buffer: Buffer A.
Elution buffer²: Tris.HCl 20mM pH 7.4.
Regeneration buffer: NaOH 0.1M.
¹ The sample can be also the ascites fluid or culture medium or egg yolk extract.
² The elution can be also realized with a buffer of your choice.

• IgG, IgA and IgM are retained at F_c region by T-GEL.
• IgG is more retained than IgA and IgM, by T-GEL.
• F_ab and F_ab' are not retained by T-GEL³.

APPLICATION

Coating conditions:
- 500 µl per tube.
- Wash 3 times.
Pre-treatment of coated tubes: Dried at room temperature for 24 hours.
Series 1: With coated tubes stored at 4°C.
Series 2: With coated tubes incubated at 37°C for 7 days.
Testing conditions:
- 50 µl of known or unknown sample + 100 µl ¹²⁵I MAb anti hCG.
- Shake 1 hour at room temperature.
- Wash 2 times.