Methods and apparatus allowing sequential chemical reactions

• US 5,516,698 A
• Filed: 04/30/1992
• Issued: 05/14/1996
• Est. Priority Date: 04/11/1990
• Status: Expired due to Fees

• Current Assignee: Ludwig Institute for Cancer Research, Ltd.
• Original Assignee: Ludwig Institute for Cancer Research, Ltd.
• Inventors: Begg, Geoffrey S., Simpson, Richard J., Burgess, Antony W.
• Primary Examiner(s): Snay, Jeffrey R.
• Application Number:
  US07/842,363
• Time in Patent Office: 1,475 Days
• Field of Search: 436/86-90, 436/149, 436/150, 436/151, 436/161, 436/173, 436/175, 436/177, 436/178, 436/180, 422/101, 422/82.01, 422/82.02, 422/186.04, 422/67, 204/299 R, 204/302, 204/180.1, 204/186
• US Class Current: 436/89
• CPC Class Codes: B01J 19/0046 Sequential or parallel reactions, e.g. for the synthesis of polypeptides or polynucleotides; Apparatus and devices for combinatorial chemistry or for making molecular arrays synthesis methods per se C40B50/00
• CPC Class Codes: B01J 2219/0059 Sequential processes
• CPC Class Codes: B01J 2219/00596 Solid-phase processes
• CPC Class Codes: B01J 2219/00689 using computers
• CPC Class Codes: B01J 2219/00698 Measurement and control of process parameters
• CPC Class Codes: B01J 2219/00725 Peptides
• CPC Class Codes: C07K 1/128 sequencing
• CPC Class Codes: C40B 40/10 Libraries containing peptides or polypeptides, or derivatives thereof
• CPC Class Codes: G01N 27/44 using electrolysis to generate a reagent, e.g. for titration
• CPC Class Codes: G01N 27/44726 using specific dyes, markers or binding molecules
• CPC Class Codes: G01N 27/4473 by electric means
• CPC Class Codes: G01N 30/06 Preparation
• CPC Class Codes: G01N 31/00 Investigating or analysing non-biological materials by the use of the chemical methods specified in the subgroup; Apparatus specially adapted for such methods
• CPC Class Codes: G01N 33/6824 involving N-terminal degradation, e.g. Edman degradation
• CPC Class Codes: Y10T 436/2575 Volumetric liquid transfer

Abstract

A first reactant is immobilized i.e. in a porous matrix (50), adjacent a sample electrode (46) within a reaction chamber. Energizing of the electrode (46) electrophoretically attracts a mobile second reactant and/or electrolytically induces appropriate reaction conditions to enhance reaction of the first and second reactants. Polarity reversals between the sample electrode (46) and remote electrodes (38), (42), (44) cause unreacted second reactant and/or by-products to migrate away from the immobilized first reactant. The techniques are useful for sequential chemical reactions such as sequencing or construction of proteins, polysaccharides and nucleic acids where cyclical additions and removals of reactants are required. The techniques are amenable to automated micro and nano scale construction and operation and allow direct electrophoretic (38) interfacing with chromatographic, HPCE and mass spectrophotometric equipment.

Claims

1. A method for conducting a chemical reaction, wherein said method comprises, immobilizing a first reactant at the reaction locus, introducing an additional reactant to the reaction locus, allowing a reaction between the first and second reactants to produce a reaction by-product in an anhydrous medium, if the by-product is not charged, imparting an electric charge to the reaction by-product and electrophoretically removing the reaction by-product from the reaction locus, and further comprising an electrolytic activation step comprising electrolytically activating a reaction between the first and additional reactants and removing remaining additional reactant and/or a reaction by-product from the reaction locus.

2. A method for conducting a chemical reaction, wherein said method comprises, immobilizing a first reactant at a reaction locus, introducing an additional reactant to the reaction locus, allowing a reaction between the first and second reactants to produce a reaction by-product in an anhydrous medium, if the by-product is not charged, imparting an electric charge to the reaction by-product, and electrophoretically removing the reaction by-product from the reaction locus, and wherein the by-product immediately after the reaction is not charged and wherein a charged surfactant is introduced to the reactant locus to impart a charge to the by-product.

3. A method of conducting a chemical reaction, the method comprising, immobilizing a first reactant at a reaction locus, electrophoretically introducing to the reaction locus at least one charged, additional reactant in an anhydrous medium, and after the reaction between the first and additional reactants removing the remaining additional reactant and/or a reaction by-product of the reaction from the reaction locus, and wherein additional reactant remaining from the reaction is electrophoretically removed from the reaction locus by energizing an electrode remote from the reaction locus with an electric charge of greater magnitude than that of the reaction locus, and further wherein the polarity of the electrode adjacent the reaction locus is reversed to enable the remote electrode to bear the greater magnitude of electric charge.

4. A method of conducting a chemical reaction, the method comprising immobilizing a first reactant at a reaction locus, electrophoretically introducing to the reaction locus at least one charged additional reactant in an anhydrous medium, and after reaction between the first and additional reactants removing the remaining additional reactant and/or a reaction by-product of the reaction from the reaction locus, and wherein additional reactant remaining from the reaction is electrophoretically removed from the reaction locus by energizing an electrode remote from the reaction locus with an electric charge of greater magnitude than that of the reaction locus, and further wherein the remote electrode defines a storage locus for additional reactant and wherein additional reactant can be electrophoretically reintroduced to the reaction locus.