By K. Dane Wittrup, Gregory L. Verdine
ISBN-10: 0124160395
ISBN-13: 9780124160392
This quantity of equipment in Enzymology seems to be at Protein Engineering for Therapeutics. The chapters provide an precious source for lecturers, researchers and scholars alike. With a global board of authors, this quantity is divided into sections that conceal topics similar to Antibodies, Protein conjugates, Peptides, Enzymes and Scaffolds
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Additional info for Protein Engineering for Therapeutics, Part A
Sample text
Transfer the supernatant to a fresh tube and add 1/5 volume of PEG/ NaCl solution to precipitate the phage. Incubate 5 min at room temperature. 11. Centrifuge for 10 min at 10,000 rpm and 4 C in a GSA rotor (16,000Âg). Decant the supernatant. Spin briefly and remove the remaining supernatant with a pipette. 12. Resuspend the phage pellet in 20 ml of PBT buffer. 13. Pellet insoluble matter by centrifuging for 5 min at 15,000 rpm and 4 C in an SS-34 rotor (27,000Âg). Transfer the supernatant to a clean tube.
0 h and transfer 100 ml to Maxisorp immunoplates coated with antigen and blocked as described previously. 13. Incubate for 15 min with gentle shaking. 14. 1. 15. Plot the OD450 reading as a function of the antigen concentration and determine the IC50 value by standard curve fitting. The IC50 is defined as the concentration of antigen that blocks 50% of antibody-phage binding to immobilized antigen. 5. DNA sequencing Once candidate clones have been identified, the sequence of the displayed antibody can be deduced by sequencing the encoding DNA encapsulated within the phage particle.
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Protein Engineering for Therapeutics, Part A by K. Dane Wittrup, Gregory L. Verdine
by Anthony
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