Purification of lectins by biospecific affinity chromatography

Area of Science:

• Biochemistry
• Affinity Chromatography

Background:

• Lectins are proteins with specific carbohydrate-binding properties.
• Purification of lectins is crucial for various biological and diagnostic applications.
• Existing methods for lectin purification can be complex or inefficient.

Purpose of the Study:

• To develop a straightforward method for preparing biospecific adsorbents for lectin purification.
• To immobilize lectin-specific sugars onto Epoxy-activated Sepharose 6 B.
• To evaluate the efficiency of these adsorbents for purifying specific lectins.

Main Methods:

• One-step reaction between Epoxy-activated Sepharose 6 B and lectin-specific sugars (N-acetyl-D-galactosamine and N-acetyl-D-glucosamine).
• Immobilization of sugars onto the Sepharose matrix.
• Application of the resulting Sepharose derivatives for lectin purification.
• Electrophoresis in polyacrylamide gels for purity assessment.

Main Results:

• Successfully prepared biospecific adsorbents by immobilizing N-acetyl-D-galactosamine and N-acetyl-D-glucosamine.
• Purified soybean lectin with a maximal capacity of 15 mg/ml gel, yielding 145 mg/100g soybeans.
• Purified wheat germ lectin with a capacity of 10 mg/ml gel, yielding 65 mg/100g wheat germ.
• Purified lectins demonstrated homogeneity via electrophoresis.

Conclusions:

• Biospecific adsorbents offer an easy and effective approach for lectin purification.
• The developed method provides high yields of purified lectins from plant sources.
• This technique is valuable for obtaining homogeneous lectins for research and applications.