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Enhancement of biological reactions on cell surfaces via macromolecular crowding

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  • Rafi Chapanian

    (Centre for Blood Research, University of British Columbia, 2350 Health Sciences Mall, Life Sciences Centre, Vancouver, British Columbia, Canada V6T 1Z3
    University of British Columbia, 2350 Health Sciences Mall, Life Sciences Centre, Vancouver, British Columbia, Canada V6T 1Z3)

  • David H. Kwan

    (University of British Columbia, 2036 Main Mall, Vancouver, British Columbia, Canada V6T 1Z1
    Centre for High-Throughput Biology, University of British Columbia)

  • Iren Constantinescu

    (Centre for Blood Research, University of British Columbia, 2350 Health Sciences Mall, Life Sciences Centre, Vancouver, British Columbia, Canada V6T 1Z3)

  • Fathima A. Shaikh

    (University of British Columbia, 2036 Main Mall, Vancouver, British Columbia, Canada V6T 1Z1)

  • Nicholas A..A. Rossi

    (Centre for Blood Research, University of British Columbia, 2350 Health Sciences Mall, Life Sciences Centre, Vancouver, British Columbia, Canada V6T 1Z3)

  • Stephen G Withers

    (University of British Columbia, 2036 Main Mall, Vancouver, British Columbia, Canada V6T 1Z1
    Centre for High-Throughput Biology, University of British Columbia
    University of British Columbia)

  • Jayachandran N Kizhakkedathu

    (Centre for Blood Research, University of British Columbia, 2350 Health Sciences Mall, Life Sciences Centre, Vancouver, British Columbia, Canada V6T 1Z3
    University of British Columbia, 2350 Health Sciences Mall, Life Sciences Centre, Vancouver, British Columbia, Canada V6T 1Z3
    University of British Columbia, 2036 Main Mall, Vancouver, British Columbia, Canada V6T 1Z1)

Abstract

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.

Suggested Citation

  • Rafi Chapanian & David H. Kwan & Iren Constantinescu & Fathima A. Shaikh & Nicholas A..A. Rossi & Stephen G Withers & Jayachandran N Kizhakkedathu, 2014. "Enhancement of biological reactions on cell surfaces via macromolecular crowding," Nature Communications, Nature, vol. 5(1), pages 1-12, December.
    • Handle: RePEc:nat:natcom:v:5:y:2014:i:1:d:10.1038_ncomms5683
    DOI: 10.1038/ncomms5683
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    Cited by:

    1. Jorik Waeterschoot & Willemien Gosselé & Špela Lemež & Xavier Casadevall i Solvas, 2024. "Artificial cells for in vivo biomedical applications through red blood cell biomimicry," Nature Communications, Nature, vol. 15(1), pages 1-17, December.

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