Affinity tuning of antibodies using yeast surface display
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Affinity tuning of antibodies using yeast surface display. Yeast surface display of I-Ani1: Two epitope tags, One “antigen” binding site. Yeast surface display of I-Ani1: single bp resolution of binding specificity. 1 bp change. 2 bp change. 3 bp change. 1 bp change. 2 bp change.

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Yeast surface display of I-Ani1: single bp resolution of binding specificity

1 bp change

2 bp change

3 bp change

1 bp change

2 bp change


Strategies for generating variants & libraries binding specificity

Rational & Computational Design

Combinatorial Libraries & Gene Shuffling

Error Prone PCR

Jim, Summer, Justin, Audrey, Umut , and others?


Optimal specific S/N ratio at physiological osmolarity binding specificity

75 mM KCl

100 mM KCl

125 mM KCl

150 mM KCl

175 mM KCl

200 mM KCl

222 mOsm

262 mOsm

316 mOsm

341 mOsm

405 mOsm

472 mOsm



JH&ST-Ani combinatorial Lib1.0 T29R E31L R72S

-ve MACS Fraction

+ve MACS Fraction



Yeast surface display of I-Ani1: ionic effects on HE specificity

ReoAni

STS3/4 Lib1.0.A2

Kd = 65nM

Kd = 20 nM

Kd = 8 nM

Standard conditions: 150mM KCl, 10mM NaCl, 5mM CaCl2, 10mM HEPES, 0.2% BSA, 0.1% Galactose, pH 7.5


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