Solid-phase Immobilization of Microbial Transglutaminase Mtg on Microbeads for Protein Conjugation

Simple SummaryContent extracted from patent full text and abstract with AI.

This invention describes a method for site-specific modification of proteins by immobilizing microbial transglutaminase (MTG) onto polymer-coated microbeads. By anchoring MTG on solid microbeads, it enables efficient, selective, and reusable protein conjugation, attaching functional molecules (like dyes, drugs, or labels) to therapeutic proteins (e.g., antibodies, antibody fragments) with high precision under physiological conditions. The immobilized enzyme allows for flow-through or batch processes, enhances selectivity for target residues, and minimizes enzyme loss during repeated uses.

Use CasesContent extracted from patent full text and abstract with AI.

• Production of antibody-drug conjugates for targeted cancer therapies.
• Labeling therapeutic proteins or antibodies with fluorescent dyes for research or diagnostics.
• Attaching metal chelators or radioactive labels to proteins for imaging or therapeutic purposes.
• Creating site-specific bioconjugates in vaccine development.
• Functionalizing proteins with cell-cytotoxic agents for immunotherapy.
• Batch processing or continuous manufacturing of modified biologics in biotech industry.
• Protein engineering for research applications, including site-specific labeling or loading of proteins with diverse functional groups.

BenefitsContent extracted from patent full text and abstract with AI.

• Achieves high site-specificity, reducing unwanted modifications and product heterogeneity.
• Allows for easy recovery and repeated reuse of the immobilized enzyme, saving costs and resources.
• Minimizes loss of enzyme and prevents contamination of product with free enzyme ('no column bleeding').
• Enables controlled, efficient, and scalable conjugation under mild, physiological conditions.
• Enhanced selectivity for specific amino acid residues, even when multiple reactive sites are present.
• Adaptable to various proteins (antibodies, fragments, peptides) and functional molecules (dyes, drugs, metal chelators, etc.).
• Suitable for automated, continuous processes (flow reactors, spin-columns) and scalable manufacturing.
• Improves product consistency and purity, which is critical for therapeutic and diagnostic use.

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Inventors & Applicants

Patent Abstract

Site-specific modification of proteins with microbial transglutaminase (MTG) is a powerful and versatile strategy for a controlled modification of proteins under physiological conditions. We present evidence that solid-phase microbead-immobilization can be used to site-specifically and efficiently attach different functional molecules important for further downstream applications to proteins of therapeutic relevance including scFV, Fab-fragment and antibodies. We demonstrate that MTG remained firmly immobilized with no detectable column bleeding and that enzyme activity was sustained during continuous operation, which allowed for a convenient recycling of the enzyme, thus going beyond solution-phase MTG conjugation. In addition it is showed that immobilized MTG shows enhanced selectivity towards a certain residue in the presence of several reactive residues which are all targeted if the conjugation was carried out in solution.