Abstract：
:In cell affinity chromatography, type-specific cell separation is based on the interaction between cell-surface receptors and an immobilized ligand on a stationary matrix. This protocol describes the preparation of monolithic polyacrylamide and polydimethylacrylamide cryogel affinity matrices that can be used as a generic type-specific cell separation approach. The supermacroporous monolithic cryogel has highly interconnected large pores (up to 100 μm) for convective migration of large particles such as mammalian cells. In this protocol, they are functionalized to immobilize a protein A ligand by a two-step derivatization of epoxy-containing cryogel monolith (reaction with ethylenediamine and glutaraldehyde). Target cells were labeled with specific antibodies and then they were captured in the cryogel through affinity with protein A. These specifically captured cells were recovered in high yields while retaining their viability by mechanical squeezing of the spongy and elastic cryogel matrices. The suggested cell separation protocol takes < 30 min for complete separation on a preprepared protein A-immobilized cryogel column.
journal_name
Nat Protocjournal_title
Nature protocolsauthors
Kumar A,Srivastava Adoi
10.1038/nprot.2010.135subject
Has Abstractpub_date
2010-11-01 00:00:00pages
1737-47issue
11eissn
1754-2189issn
1750-2799pii
nprot.2010.135journal_volume
5pub_type
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