Peptide or protein sequencing method and apparatus
AssigneeDurrum Instrument Corporation
InventorWilliam J. Dreyer
Life of this patent
Estimating the term…
Abstract
A method and apparatus for the sequential degradation of protein or peptide molecules by successive coupling and cleavage reactions. Such molecules are immobilized on a macroporous reaction support surface and placed in a flowthrough reaction chamber which is mounted in a sequencer. In the Edman sequencing technique, reagents driven by pressurized inert gas are passed through the reaction chamber as follows: (a) liquid or vapor coupling reagent, (b) coupling base vapor, (c) washing solvent, (d) inert gas to partially dry the support surface, (e) cleavage reagent vapor to cleave amino acid derivatives from the immobilized coupled protein or peptide chains, (f) liquid extracting solvent to withdraw the cleaved amino acid derivative. An automated sequencer to carry out said method.
Claims29
In a method for the sequential degradation of protein or peptide chains in a reaction chamber by successive coupling and cleavage reactions, the steps of:
- a. immobilizing said protein or peptide on a macroporous reaction support surface disposed in the reaction chamber, said immobilization being performed either by nonchemical sorptive deposition onto said support surface or by chemical linkage to said support surface,
- b. passing coupling reagent through the reaction chamber to contact said immobilized protein or peptide,
- c. directing a pressurized vapor stream comprising coupling base vapor through the reaction chamber while said coupling reagent is in contact with said protein or peptide to provide basic environment for coupling of said coupling reagent to said chains,
- d. flowing a liquid washing solvent through the reaction chamber to remove unreacted coupling reagent and other contaminants from the support surface,
- e. directing a pressurized stream of inert carrier gas through the reaction chamber after step (d) to at least partially dry said support surface,
- f. directing a pressurized vapor stream comprising cleavage reagent vapor through the reaction chamber for sufficient time to cleave amino acid derivatives from said coupled protein or peptide, and
- g. flowing a liquid extracting solvent through the reaction chamber after step (f) to extract and withdraw said cleaved amino acid derivative.
In a method for the sequential degradation of protein or peptide chains in a reaction chamber by successive coupling and cleavage reactions, the steps of:
- a. immobilizing the protein or peptide on a macroporous reaction support surface disposed in the reaction chamber, said immobilization being performed either by nonchemical sorptive deposition onto said support surface or by chemical linkage to said support surface,
- b. passing coupling reagent in liquid form through the reaction chamber to contact and deposit on said immobilized protein or peptide chains, and
- c. directing a pressurized vapor stream comprising coupling base vapor through the reaction chamber while said protein or peptide chains are wet with coupling reagent liquid to provide a basic environment for coupling of said coupling reagent to said immobilized protein or peptide.
In a method for the sequential degradation of protein or peptide chains in a reaction chamber by successive coupling and cleavage reactions, the steps of:
- a. immobilizing said peptide or protein chains on a macroporous reaction support surface disposed in the reaction chamber, said immobilization being performed either by nonchemical sorptive deposition onto said support surface or by chemical linkage to said support surface,
- b. chemically coupling a coupling reagent to said protein or peptide chains,
- c. passing a pressurized vapor stream comprising cleavage reagent vapor through the reaction chamber for sufficient time to cleave amino acid derivatives from said coupled protein or peptide chains, and
- d. flowing a liquid extracting solvent through the reaction chamber to extract and withdraw said cleaved amino acid derivatives.
In a method for the sequential degradation of protein or peptide chains in a reaction chamber by successive coupling and cleavage reactions, the steps of:
- a. immobilizing said protein or peptide chains on a macroporous surface in said chamber by nonchemical sorptive deposition,
- b. chemically coupling a coupling reagent to said protein or peptide chains in the presence of a coupling base, and
- c. directing a cleavage reagent through the reaction chamber for sufficient time to cleave amino acid derivative from said coupled protein or peptide chains, and
- d. flowing a liquid extracting solvent through the reaction chamber to extract and withdraw said cleaved amino acid derivative.
Dependencies are inferred from the claim text.