Phage Display: Simple Evolution in a Petri Dish George P. Smith - - PowerPoint PPT Presentation

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Phage Display: Simple Evolution in a Petri Dish George P. Smith - - PowerPoint PPT Presentation

Sep 11, 2022 273 likes •718 views

Phage Display: Simple Evolution in a Petri Dish George P. Smith Division of Biological Sciences University of Missouri Nobel Prize Lecture in Chemistry December 8, 2018 Margie did not invent phage display My science community

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Phage Display: Simple Evolution in a Petri Dish

George P. Smith Division of Biological Sciences University of Missouri Nobel Prize Lecture in Chemistry December 8, 2018

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Margie did not invent phage display

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My science community

  • Molecular biologists
  • Evolutionary biologists
  • Immunologists
  • Protein chemists
  • Phage biologists
  • Mathematicians
  • Philosophers of science
  • Bayesian statisticians

Phage display

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Our science community

  • Molecular biologists
  • Evolutionary biologists
  • Immunologists
  • Protein chemists
  • Phage biologists
  • Mathematicians
  • Philosophers of science
  • Bayesian statisticians

Phage display

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Phage = virus that infects bacteria

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Filamentous phage

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Filamentous phage

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Filamentous phage ~1 µm 6 nm

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Crash course in molecular biology

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Crash course in molecular biology

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Filamentous phage infection cycle (simplified)

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Bob Webster, Department of Biochemistry, Duke University (now retired to North Carolina coast)

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Paul Modrich Nobel Prize in Chemistry, 2015

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Why is that interesting? Please wait…

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Darn good colleague Steve Parmley

Steve developed a practical phage display vector (after some false starts like pIG3C) and affinity selection as grad student, 1985-1988.

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Jamie Scott first demonstrated affinity selection

  • f peptides from

large random peptide libraries as postdoc, 1988-1991. Darn good colleague Jamie Scott

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Robert Davis came to the lab as chief manager and technician in the summer of 1989. We calculate that he sequenced a million DNA bases using old-fashioned radioactive technology. Darn good colleague Robert Davis

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Fred Richards at Yale, late 50’s; brought to our lab by John Ladbury and David Schultz

The S-protein system

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“

Pretend “receptor” Pretend “natural ligand”

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S-protein “receptor”

Jinan Yu, now a researcher at Hainan University, China.

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Random peptide library

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Random peptide library

1015 phages representing 250 million phage clones, each clone displaying a different 15-amino acid guest peptide. [T. Nishi et al., FEBS Letters 399, 237–240 (1996)]

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?

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Affinity selection

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NRAWSEFLWQHLAPV KETAAAKFERQHMDSSTSAA

Selected peptide

Dominant sequence among selected peptides

Buried amino acids

(one-letter abbreviations for amino acids)

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NRAWSEFLWQHLAPV KETAAAKFERQHMDSSTSAA

Selected peptide S-peptide

Dominant sequence among selected peptides aligns with S-peptide “natural ligand”

Buried amino acids

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Evolution in the living world

  • Diversification
  • Natural selection
  • Adaptation

Affinity-selection from random peptide libraries

  • Construction of library
  • Affinity selection
  • Peptide with desired

activity

Artificial evolution in the petri dish

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Cure for S- protein disease

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Cure for S- protein disease Marvel 2 Selection scheme 2

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Marvel 2 Marvel 3 Selection scheme 2 Cure for S- protein disease