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Non-robotic high-throughput setup for manual assembly of nanolitre vapour-diffusion protein crystallization screens
Nanolitre‐sized drops are characteristic of high‐throughput protein crystallization screening. Traditionally, reliable nanolitre drop dispensing has required the use of robotics. This work describes the design and development of a protocol for the reproducible manual assembly of nanolitre‐sized prot...
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Published in: | Journal of applied crystallography 2012-10, Vol.45 (5), p.1061-1065 |
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Main Authors: | , , |
Format: | Article |
Language: | English |
Subjects: | |
Citations: | Items that this one cites Items that cite this one |
Online Access: | Get full text |
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Summary: | Nanolitre‐sized drops are characteristic of high‐throughput protein crystallization screening. Traditionally, reliable nanolitre drop dispensing has required the use of robotics. This work describes the design and development of a protocol for the reproducible manual assembly of nanolitre‐sized protein vapour‐diffusion crystallization trials in a 96/192‐drop format. The protocol exploits the repetitive‐pipetting mode of handheld motorized pipettes together with simple tools available in standard laboratories. The method saves precious protein material without sacrificing the effectiveness of the screening process. To verify the approach, two monoclonal antibody Fab fragments were crystallized alone and in a complex with tau peptide antigens in 0.2–0.5 µl drops. Crystals grown directly from the screen conditions in sitting drops on 96‐well plates diffracted up to 1.6 Å resolution on a synchrotron source. The results proved that successful crystallization in nanolitre high‐throughput format is affordable even in the absence of expensive robotic instrumentation. |
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ISSN: | 1600-5767 0021-8898 1600-5767 |
DOI: | 10.1107/S0021889812036527 |