Composite imprinted macroporous hydrogels for haemoglobin purification from cell homogenate

Area of Science:

• Biochemistry
• Materials Science
• Biotechnology

Background:

• Haemoglobin (Hb) purification is crucial for its therapeutic applications as an oxygen carrier.
• Existing purification methods face challenges in selectivity and efficiency.
• Development of advanced chromatographic techniques is needed for effective Hb isolation.

Purpose of the Study:

• To develop and evaluate novel affinity chromatography columns for efficient haemoglobin purification.
• To compare two distinct molecular imprinting strategies for creating Hb-selective adsorbents.
• To assess the performance of these columns using non-clarified biological samples.

Main Methods:

• Preparation of affinity adsorbents using molecular imprinting techniques.
• Immobilization of imprinted particles onto cryogel supports via Pickering emulsion polymerization and covalent bonding.
• Direct protein imprinting of haemoglobin onto acrylamide cryogel matrices.
• Evaluation of dynamic binding capacity, selectivity, and performance in batch mode.

Main Results:

• Two types of affinity chromatography columns were successfully developed using molecular imprinting.
• The composite cryogel with immobilized particles exhibited a dynamic binding capacity of 5.2 mg/g.
• The directly imprinted acrylamide cryogel showed a dynamic binding capacity of 3.6 mg/g.
• Both columns demonstrated high selectivity for haemoglobin, even in the presence of interfering substances.

Conclusions:

• Molecularly imprinted cryogels offer a robust and selective platform for haemoglobin purification.
• The developed affinity chromatography columns show potential for clinical applications requiring high-purity haemoglobin.
• Protein imprinting directly onto cryogels presents an efficient alternative for adsorbent preparation.