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Affinity tuning of antibodies using yeast surface display

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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Affinity tuning of antibodies using yeast surface display

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  1. Affinity tuning of antibodies using yeast surface display

  2. Yeast surface display of I-Ani1: Two epitope tags, One “antigen” binding site

  3. 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

  4. Strategies for generating variants & libraries Rational & Computational Design Combinatorial Libraries & Gene Shuffling Error Prone PCR Jim, Summer, Justin, Audrey, Umut , and others?

  5. Optimal specific S/N ratio at physiological osmolarity 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

  6. Analysis of computationally designed I-Ani1 variant: K24N T29R E31L R72S

  7. JH&ST-Ani combinatorial Lib1.0 -ve MACS Fraction +ve MACS Fraction

  8. Targeting STS3 and STS4 to generate a high-affinity I-Ani1 variant

  9. 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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