Enhancement of biological reactions on cell surfaces via macromolecular crowding

The reaction of macromolecules such as enzymes and antibodies with cell surfaces is often an inefficient process, requiring large amounts of expensive reagent. Here we report a general method based on macromolecular crowding with a range of neutral polymers to enhance such reactions, using red blood...

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Bibliographic Details
Published in:Nature communications 2014-08, Vol.5 (1), p.4683-4683, Article 4683
Main Authors: Chapanian, Rafi, Kwan, David H., Constantinescu, Iren, Shaikh, Fathima A., Rossi, Nicholas A..A., Withers, Stephen G, Kizhakkedathu, Jayachandran N
Format: Article
Language:English
Subjects:
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ABO Blood-Group System - chemistry
Dextrans - chemistry
Diffusion
Erythrocytes - chemistry
Erythrocytes - cytology
Excipients - chemistry
Ficoll - chemistry
Fluorescence Recovery After Photobleaching
Genes, Reporter
Glycerol - chemistry
Glycoside Hydrolases - chemistry
Humanities and Social Sciences
Humans
Hydrolysis
Microscopy, Confocal
multidisciplinary
Polymers - chemistry
Povidone - chemistry
Recombinant Fusion Proteins - chemistry
Science
Science (multidisciplinary)
Surface Properties
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Summary:The reaction of macromolecules such as enzymes and antibodies with cell surfaces is often an inefficient process, requiring large amounts of expensive reagent. Here we report a general method based on macromolecular crowding with a range of neutral polymers to enhance such reactions, using red blood cells (RBCs) as a model system. Rates of conversion of type A and B red blood cells to universal O type by removal of antigenic carbohydrates with selective glycosidases are increased up to 400-fold in the presence of crowders. Similar enhancements are seen for antibody binding. We further explore the factors underlying these enhancements using confocal microscopy and fluorescent recovery after bleaching (FRAP) techniques with various fluorescent protein fusion partners. Increased cell-surface concentration due to volume exclusion, along with two-dimensionally confined diffusion of enzymes close to the cell surface, appear to be the major contributing factors. Optimizing cell-surface biological reactions is an important goal of biotechnology and industrial processes. Here the authors use macromolecular crowding to enhance the enzymatic conversion of red blood cells to the universal type O blood type, using orders of magnitude less enzyme than was previously required.
ISSN:2041-1723
2041-1723
DOI:10.1038/ncomms5683