Method of extraction of an enzyme from plant or animal tissue
AssigneeSyngenta Participations AG
InventorMahathelge Dilip Dias
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Abstract
This invention provides a method for the extraction and detection of a peptide from transgenic plant tissues wherein a non-immunogenic solubility-promoting compound is used to release the enzyme into the solution fraction during the purification process. In some embodiments, this invention provides a method for the extraction and detection of the enzyme Amy797E, which is a heterologous thermo-tolerant α-amylase, from the tissues of corn event 3272 using a non-immunogenic amylase during the purification process. This invention allows for a limit of detection of 1:1000 of Amy797E in an enzyme-linked immunosorbent assay (ELISA).
Claims13
A method for extraction of a polypeptide bound to an insoluble substrate within a biological sample, comprising:
- a) homogenizing the biological sample in the presence of extraction buffer and a solubility-promoting compound, wherein the solubility-promoting compound is a polypeptide;
- b) incubating the sample to permit solubilization of the polypeptide;
- c) centrifuging the sample to separate the soluble and insoluble fractions, wherein the polypeptide fractionates into the soluble fraction.
A method for extraction of a polypeptide bound to an insoluble substrate within plant tissue, comprising:
- a) homogenizing the plant tissue sample in the presence of extraction buffer and a non-immunoreactive, solubility-promoting compound, wherein the solubility-promoting compound is a polypeptide;
- b) incubating the sample to permit solubilization of the polypeptide;
- c) centrifuging the sample to separate the soluble and insoluble fractions, wherein the polypeptide fractionates into the soluble fraction.
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Dependencies are inferred from the claim text.