Match the type of chromatographic technique (List I) with the protein property (List II)
LIST I                                                                                                      LIST II
A. Dye affinity                                                                                       I. Charge and isoelectric point
B. Chromato focussing                                                                       II. Molecular size
C. Covalent                                                                                           III. Structure and hydrophobicity
D. Gel Filtration                                                                                  IV. Thiol groups
Choose the correct answer from the options given below:
1. A-III, B-I, C-IV, D-II
2. A-III, B-II, C-IV, D-I
3. A- II. B-III, C-I, D-IV
4. A-I, B-IV, C-II, D-III

Chromatographic Techniques and Their Role in Protein Separation

Introduction

Chromatography is a powerful technique used for protein purification and separation based on various properties such as size, charge, affinity, and hydrophobicity. Different types of chromatography exploit these characteristics to isolate proteins for biochemical and pharmaceutical research.

In this article, we will match chromatographic techniques with the corresponding protein properties and discuss their significance in biotechnology and molecular biology.

Correct Answer: Option 1: A-III, B-I, C-IV, D-II

Keyphrase: Chromatographic Techniques in Protein Purification

1. Dye Affinity Chromatography

• Principle: Proteins interact with immobilized dye ligands based on their hydrophobic regions and structure.

• Mechanism:

  • Dyes mimic cofactors or substrates of enzymes.

  • Proteins with similar binding sites interact with the dye matrix.

• Example: Cibacron Blue dye is commonly used to purify dehydrogenases and kinases.

• Applications:

  • Separation of enzymes, albumins, and binding proteins.

  • Used in biopharmaceutical industries.

2. Chromatofocusing

• Principle: Proteins are separated based on charge and isoelectric point (pI).

• Mechanism:

  • A pH gradient is established using buffering solutions.

  • Proteins migrate to their isoelectric point (pI), where they carry no net charge and are eluted.

• Example: Used to separate monoclonal antibodies and recombinant proteins.

• Applications:

  • Purification of charge-based protein variants.

  • Used in pharmaceutical and clinical research.

3. Covalent Chromatography

• Principle: Proteins containing thiol (-SH) groups form covalent bonds with reactive matrix ligands.

• Mechanism:

  • The chromatography matrix is functionalized with maleimide or iodoacetate groups.

  • Proteins with free cysteine residues (thiol groups) bind covalently.

• Example: Used to purify enzymes with active thiol groups.

• Applications:

  • Separation of enzymes and proteins with cysteine residues.

  • Used in bioconjugation and drug development.

4. Gel Filtration Chromatography (Size Exclusion Chromatography)

• Principle: Proteins are separated based on molecular size.

• Mechanism:

  • The column contains porous beads that allow smaller molecules to enter, delaying their elution.

  • Larger molecules move faster and elute earlier.

• Example: Used for purifying antibodies, enzymes, and protein complexes.

• Applications:

  • Determination of molecular weight and protein folding.

  • Used in biopharmaceutical and structural biology research.

Applications of Chromatographic Techniques

1. Protein Purification

   • Essential for biotechnology, pharmaceutical, and research laboratories.

   • Used for purifying therapeutic proteins, enzymes, and vaccines.

2. Clinical Diagnostics

   • Used in blood serum analysis and biomarker detection.

   • Important for diagnosing cancer, infections, and metabolic disorders.

3. Drug Discovery and Development

   • Identifies protein-drug interactions.

   • Used for purifying monoclonal antibodies and recombinant proteins.

4. Food and Agricultural Research

   • Detection of protein contaminants in food.

   • Analysis of nutritional content and food safety.

Conclusion

Chromatography is a crucial technique in biological and pharmaceutical research. Each type of chromatography targets specific protein properties, ensuring high-purity protein separation. Understanding these methods is essential for biochemists, molecular biologists, and biotechnologists.

For more insights into chromatography techniques, CSIR NET, GATE, IIT JAM, DBT BET JRF preparations, visit Let’s Talk Academy.