Methods for the production of proteins with a desired function
First Claim
1. A method for producing an antibody, antibody variable region, or a portion thereof which has an antibody binding region with a desired function, comprising:
- (a) obtaining from a warm-blooded animal a population of antibody-forming B cells or antibody-forming progeny thereof wherein the B cells or progency thereof are non-hybridoma or non-Epstein Barr virus (EBV) transformed, and wherein the B cells or progency thereof contain at least one cell that produces an antibody exhibiting a desired function, followed by;
(b) suspending the population of antibody-forming B cells or progency thereof in a medium, the medium having an indicator system incorporated therein which indicates the presence and location of a cell which forms antibodies exhibiting the desired function;
(c) identifying a cell forming an antibody exhibiting the desired function;
(d) isolating the identified antibody-forming cell from the medium;
(e) determining the amino acid sequence of the variable region or a portion thereof which exhibits the desired function of the antibody produced by the isolated antibody-forming cell; and
(f) synthesizing an antibody, antibody variable region, or a portion thereof which has the amino acid sequence of the variable region or portion thereof which exhibits the desired function.
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Accused Products
Abstract
The present invention provides a method for producing proteins with a desired function, generally comprising the steps of (a) providing a population of antibody-forming cells suspected of containing at least one cell capable of producing an antibody exhibiting a desired function; (b) suspending the population of antibody-forming cells in a medium, the medium having an indicator system incorporated therein, the indicator system also being capable of indicating the presence and location of a cell which forms antibodies exhibiting the desired function; (c) identifying a cell forming an antibody exhibiting the desired function; (d) isolating the identified antibody-forming cell from the medium; (e) determining the amino acid sequence of the variable region or a portion thereof which coffers the desired function of the antibody produced by the isolated antibody-forming cell; and (f) synthesizing a protein with a desired function, the protein containing the amino acid sequence of the variable region or portion thereof which confers the desired function.
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Citations
38 Claims
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1. A method for producing an antibody, antibody variable region, or a portion thereof which has an antibody binding region with a desired function, comprising:
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(a) obtaining from a warm-blooded animal a population of antibody-forming B cells or antibody-forming progeny thereof wherein the B cells or progency thereof are non-hybridoma or non-Epstein Barr virus (EBV) transformed, and wherein the B cells or progency thereof contain at least one cell that produces an antibody exhibiting a desired function, followed by; (b) suspending the population of antibody-forming B cells or progency thereof in a medium, the medium having an indicator system incorporated therein which indicates the presence and location of a cell which forms antibodies exhibiting the desired function; (c) identifying a cell forming an antibody exhibiting the desired function; (d) isolating the identified antibody-forming cell from the medium; (e) determining the amino acid sequence of the variable region or a portion thereof which exhibits the desired function of the antibody produced by the isolated antibody-forming cell; and (f) synthesizing an antibody, antibody variable region, or a portion thereof which has the amino acid sequence of the variable region or portion thereof which exhibits the desired function. - View Dependent Claims (3, 4, 5, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38)
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2. A method for producing an antibody, antibody variable region, or a portion thereof which has an antibody binding region with a desired function, comprising:
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(a) obtaining from a warm-blooded animal a population of antibody-forming B cells or antibody-forming progeny thereof, wherein the B cells or progeny thereof are non-hybridoma or non-EBV transformed, and wherein the B cells or progeny thereof contain at least one cell that produces an antibody exhibiting a desired function, followed by; (b) suspending the population of antibody-forming B cells or progeny thereof in a medium, the medium having all indicator system incorporated therein which indicates, the presence and location of a cell which forms antibodies exhibiting the desired function; (c) identifying a cell forming an antibody exhibiting the desired function; (d) isolating the identified antibody-forming cell from the medium; (e) determining the amino acid sequence of the variable region or a portion thereof which exhibits the desired function of the antibody produced by the isolated antibody-forming cell; (f) incorporating the DNA sequence corresponding to the amino acid sequence of said variable region or portion thereof conferring the desired function into a vector which directs the expression and secretion of said antibody, antibody variable region, or a portion thereof which has an antibody binding region with a desired function; (g) transletting said vector into a host cell; (h) growing said host cells in an appropriate medium; and (i) isolating the antibody, antibody variable region, or a portion thereof which has an antibody binding region with the desired function.
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6. A method for producing an antibody, antibody variable region, or a portion thereof which has an antibody binding region with a desired function, comprising:
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(a) obtaining from a warm-blooded animal a population of antibody-forming B cells or antibody-forming progeny thereof, wherein the B cells or progeny thereof are non-hybridoma or non-EBV transformed, and wherein the B cells or progeny thereof contain at least one cell that produces an antibody exhibiting a desired function;
followed by(b) suspending the population of antibody-forming B cells or progeny thereof in a medium, the medium having an indicator system incorporated therein which indicates the presence and location of a cell which forms antibodies exhibiting the desired function; (c) identifying a cell forming an antibody exhibiting the desired function; (d) isolating the identified antibody-forming cell from the medium; (e) recovering RNA from said isolated antibody-forming cell; (f) generating cDNA from said RNA; (g) amplifying the cDNA of the variable region or a portion thereof which confers the desired function of the antibody produced by the isolated antibody-forming cell; (h) incorporating the amplified cDNA into a vector capable of directing the expression and secretion of an antibody, antibody variable region, or a portion thereof which has an antibody binding region with the desired function; (i) transfecting said vector into a host cell; (j) growing said host cells in an appropriate medium; and (k) isolating the antibody, antibody variable region, or a portion thereof which has an antibody binding region with the desired function from said host cell.
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7. A method for producing an antibody, antibody variable region, or a portion thereof which has an antibody binding region with a desired function, comprising:
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(a) obtaining from a warm-blooded animal a population of antibody-forming B cells or anti-forming progeny thereof, wherein the B cells or progeny thereof are non-hybridoma or non-EBV transformed, and wherein the B cells or progeny thereof contain at least one cell that produces an antibody exhibiting a desired function, followed by; (b) suspending the population of antibody-forming B cells or progeny thereof in a medium, the medium having an indicator system incorporated therein which indicates the presence and location of a cell which forms antibodies exhibiting the desired function; (c) identifying a cell forming an antibody exhibiting the desired function; (d) isolating the identified antibody-forming cell from the medium; (e) recovering RNA from said isolated antibody-forming cell; (f) amplifying the RNA of the variable region or a portion thereof which confers the desired function of the antibody produced by the isolated antibody-forming cell; (g) generating cDNA from said RNA; (h) incorporating the cDNA into a vector capable of directing the expression and secretion of an antibody, antibody variable region, or a portion thereof which has an antibody binding region with the desired function; (i) transfecting said vector into a host cell; (j) growing said host cells in an appropriate medium; and (k) isolating the antibody, antibody variable region, or a portion thereof which has an antibody binding region with the desired function from said host cell.
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Specification