Plane substrate packed polymer composition and its application
A polymer and coating technology, used in biochemical equipment and methods, microbial assay/inspection, instruments, etc., can solve problems such as high cost, high non-specific binding activity, and complexity
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Embodiment 1
[0057] Example 1: Optimizing the preparation of substituted polymers
[0058] (1). Preparation of DEAE-dextran / DEAE-agarose:
[0059] Dextran was purchased from Shanghai Chemical Reagent Company, agarose was purchased from Sigma Company, and DEAE was purchased from Baoding Jiahe Fine Chemical Co., Ltd.
[0060] For the preparation method, refer to an article written by one of the inventors of the present invention (COATED SILICA SUPPORTS FOR HIGH-PERFORMANCE AFFINITY CHROMATOGRAPHY OF PROTEIN, Journal of Chromatography, 476, (1989) 195-203,).
[0061] DEAE substitution rate determined by acid-base titration: 7-14%.
[0062] (2). Preparation of hydrazide-polyacrylamide:
[0063] Hydrazide-polyacrylamide was prepared according to the method of JK. Inman and HM Dintzis (Reference: The derivatization of cross-linked polyacrylamidebeads, Am. J. Epidemiol. 143: 263-268, 1969). The nonionic polyacrylamide used in this example is produced by Beijing Xitao Technology Development Co....
Embodiment 2
[0067] Embodiment 2: Preparation of polymer-coated film base
[0068] The coating polymers used in this example are shown in Table 1, and the substrates used are respectively microscopic slides (U.S. TeleChem International, Inc) used to prepare chip bases and 96-well microwells used to prepare enzyme-labeled plate bases Board (Shenzhen Jincanhua Industrial Co., Ltd.).
[0069] 1). Coating of polymer on the substrate
[0070] Dissolve the coating polymers in Table 1 (including polyvinylpyrrolidone, polysaccharide derivatives and polyacrylamide derivatives) into PBS buffer respectively to obtain a coating polymer aqueous solution with a concentration of 1% (W / V) . Then they were diluted 40-100 times respectively. The microscope slides were placed in the obtained dilution of the coated polymer, coated overnight at room temperature, washed and dried, and used after passing the quality inspection. Add 100-200 ul of the obtained coating polymer dilution to each well of a 96-well...
Embodiment 3
[0079] Embodiment 3: the preparation of chip
[0080] The coating polymer used in this example is selected from Table 1, and the sheet base used is selected from Table 2. Microscope slides as substrates and amino slides as reference photobases were purchased from TeleChem International, Inc., USA. The ligands used in this embodiment include: HCV antigen (Beijing People's Hospital Institute of Liver Diseases), HIV 1+2 Antigen (Beijing People's Hospital Institute of Liver Diseases) and EBV antigen. EBV antigen is a peptide antigen synthesized according to the method of D.Trabchand-Bunel (Detection of human antibodies using "convergent" combinatorial peptide libraries or "mixotopes" designed from anonvariable antigen: application to the EBVviral capsid antigen p18, D.Trabchand-Bunel et al., J. Peptide Res, 52.1998, pp495-508).
[0081] Before the ligand or ligand / polymer is coated, a highly hydrophobic silicone coating (Chengdu Chenguang Chemical Design Institute) is coated on...
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