Nanoliter plates—versatile tools for the screening of split-and-mix libraries on-bead and off-bead

2010 ◽  
Vol 46 (13) ◽  
pp. 2209 ◽  
Author(s):  
Grégory Upert ◽  
Christoph A. Merten ◽  
Helma Wennemers
Keyword(s):  
Split And Mix

Using Catalyst-Substrate Coimmobilization for the Discovery of Catalysts for Asymmetric Aldol Reactions in Split-and-Mix Libraries

2005 ◽  
Vol 24 (10) ◽  
pp. 1158-1163 ◽  
Author(s):  
Philipp Krattiger ◽  
Roman Kovàsy ◽  
Jefferson D. Revell ◽  
Helma Wennemers
Keyword(s):  
Aldol Reactions ◽  
Asymmetric Aldol ◽  
Split And Mix

Catalyst–Substrate Coimmobilization: A Strategy for Catalysts Discovery in Split-and-Mix Libraries

2003 ◽  
Vol 42 (15) ◽  
pp. 1722-1724 ◽  
Author(s):  
Philipp Krattiger ◽  
Catherine McCarthy ◽  
Andreas Pfaltz ◽  
Helma Wennemers
Keyword(s):  
Split And Mix

scholarly journals Split & mix assembly of DNA libraries for ultrahigh throughput on-bead screening of functional proteins

2020 ◽  
Vol 48 (11) ◽  
pp. e63-e63 ◽  
Author(s):  
Laurens Lindenburg ◽  
Tuomas Huovinen ◽  
Kayleigh van de Wiel ◽  
Michael Herger ◽  
Michael R Snaith ◽  
...  
Keyword(s):  
Amino Acid ◽  
Directed Evolution ◽  
Functional Screening ◽  
Library Synthesis ◽  
Oil Emulsion ◽  
Site Saturation ◽  
Emulsion Droplets ◽  
Dna Libraries ◽  
Screening Platform ◽  
Split And Mix

Abstract Site-saturation libraries reduce protein screening effort in directed evolution campaigns by focusing on a limited number of rationally chosen residues. However, uneven library synthesis efficiency leads to amino acid bias, remedied at high cost by expensive custom synthesis of oligonucleotides, or through use of proprietary library synthesis platforms. To address these shortcomings, we have devised a method where DNA libraries are constructed on the surface of microbeads by ligating dsDNA fragments onto growing, surface-immobilised DNA, in iterative split-and-mix cycles. This method—termed SpliMLiB for Split-and-Mix Library on Beads—was applied towards the directed evolution of an anti-IgE Affibody (ZIgE), generating a 160,000-membered, 4-site, saturation library on the surface of 8 million monoclonal beads. Deep sequencing confirmed excellent library balance (5.1% ± 0.77 per amino acid) and coverage (99.3%). As SpliMLiB beads are monoclonal, they were amenable to direct functional screening in water-in-oil emulsion droplets with cell-free expression. A FACS-based sorting of the library beads allowed recovery of hits improved in Kd over wild-type ZIgE by up to 3.5-fold, while a consensus mutant of the best hits provided a 10-fold improvement. With SpliMLiB, directed evolution workflows are accelerated by integrating high-quality DNA library generation with an ultra-high throughput protein screening platform.

Computer Aided Analysis of Split and Mix Combinatorial Libraries

2001 ◽  
Vol 3 (1) ◽  
pp. 20-27 ◽  
Author(s):  
Sid Topiol ◽  
John Davies ◽  
Sundararajan Vijayakuma ◽  
James R. Wareing
Keyword(s):  
Combinatorial Libraries ◽  
Computer Aided Analysis ◽  
Computer Aided ◽  
Split And Mix

scholarly journals Silver Nanoparticle Formation in Different Sizes Induced by Peptides Identified within Split-and-Mix Libraries

2009 ◽  
Vol 48 (20) ◽  
pp. 3661-3664 ◽  
Author(s):  
Kirsten Belser ◽  
Tünde Vig Slenters ◽  
Conelious Pfumbidzai ◽  
Grégory Upert ◽  
Laurent Mirolo ◽  
...  
Keyword(s):  
Silver Nanoparticle ◽  
Nanoparticle Formation ◽  
Split And Mix

scholarly journals Silver Nanoparticle Formation in Different Sizes Induced by Peptides Identified within Split-and-Mix Libraries

2009 ◽  
Vol 121 (20) ◽  
pp. 3715-3718 ◽  
Author(s):  
Kirsten Belser ◽  
Tünde Vig Slenters ◽  
Conelious Pfumbidzai ◽  
Grégory Upert ◽  
Laurent Mirolo ◽  
...  
Keyword(s):  
Silver Nanoparticle ◽  
Nanoparticle Formation ◽  
Split And Mix
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