Reversible method for covalent immobilization of biochemicals

Details Reversible method for covalent immobilization of biochemicals Reversible method for covalent immobilization of biochemicals

1980-06-20

1981-08-18

Schain, Howard E.

Chemistry of carbon compounds

Miscellaneous organic carbon compounds

C-metal

106308B, 106308Q, 260112B, 2601125R, 435188, 427220, C09C 308, C09C 310, C09C 312, C12N 996

Patent

active

042845530

ABSTRACT:
A method for the reversible immobilization of proteins through covalent attachment to inorganic oxide surfaces, such as glass or silica, via thioester containing coupling chains. Resulting succinamidopropyl-surfaces are converted into the acyl derivative using anhydrous thionyl chloride, and then treated with either 3-mercaptopropionic acid or mercaptoacetic acid. Thus derivatized, the material is dried, and can be stored for extended periods of time without detectable loss of reactivity with proteins. Immobilization of proteins can be readily achieved simply by suspending the derivatized material in buffer (pH=5 to 8) and recycling the protein solution through the material. Following immobilization, protein molecules can be released from the surface by cleavage of the coupling chains at the thioester linkage through treatment with hydroxylamine.

REFERENCES:
patent: 4043871 (1977-08-01), Blixt et al.
patent: 4068024 (1978-01-01), Laufer
Nature, Cuatrecasas, vol. 228, pp. 1327-1328, (1970).
Methods in Enzymology, vol. 44, 1976, pp. 516-527, (1976).
Fed. Proc., 32, p. 496, (1973), Brown et al.
Biochemistry, 15, pp. 4215-4222, Chan et al., (1976).
Biochem. Biophys. Res. Comm., 48, pp. 1068-1073, Brown et al., (1972).
Biochim-Biophysica Acta, 334, pp. 243-256, Cho et al., (1974).

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