Preanalysis treatment method for sample, and sample pretreatment system
Objects of the disclosure are to provide a preanalysis treatment method for a sample, which can reduce mixing of a reagent into the sample, and to provide a sample pretreatment system capable of performing the preanalysis treatment method. An aspect of the present embodiment is a preanalysis treatment method for a sample, which includes a step A of mixing the sample and a reagent A containing a support and a reducing agent immobilized on the support, a step B of mixing the sample and a reagent B containing a support and an enzyme immobilized on the support, a step X-A of, after the step A, separating the reagent A and a supernatant from each other, and a step X-B of, after the step B, separating the reagent B and a supernatant from each other.
1 . A preanalysis treatment method for a sample containing a peptide or a protein, comprising:
a step A of mixing the sample and a reagent A containing a support and a reducing agent immobilized on the support;
a step X-A of, after the step A, separating the reagent A and a supernatant from each other;
a step C of mixing the supernatant obtained in step X-A and a reagent C containing a support and an alkylating agent immobilized on the support;
a step X-C of, after the step C, separating the reagent C and a supernatant from each other;
a step B of mixing the supernatant obtained in step X-C and a reagent B containing a support and an enzyme immobilized on the support; and
a step X-B of, after the step B, separating the reagent B and a supernatant from each other.
2 . The preanalysis treatment method according to claim 1 , wherein
the reducing agent contained in the reagent A contains at least one thiol or phosphine group as a partial structure,
the enzyme contained in the reagent B is protease, and
the supports each have an average diameter of 0.5 μm to 500 μm.
3 . The preanalysis treatment method according to claim 1 , further comprising:
a step of heating in a range of 30° C. to 70° C.
4 . The preanalysis treatment method according to claim 1 , wherein
the reagent A contains the reducing agent in an amount of two equivalents to 1,000,000 equivalents per equivalent of disulfide bonds in the sample when added in step A,
the enzyme includes at least one enzyme selected from trypsin, chymotrypsin, pepsin, Lys-C, Lys-N, and Glu-C, and
the supports each contain at least one material selected from silica, polystyrene, acrylic resins, polysaccharides, elementary metals, semiconductors, metal oxides, and magnetic metal oxides.
5 . The preanalysis treatment method according to claim 1 , wherein
the steps X-A and X-B each include at least one method selected from natural settling, centrifugation, magnetic accumulation, sonic trapping, and filtration.