ACCELERATED HIGH-SENSITIVITY MICROSEQUENCING OF PROTEINS AND PEPTIDES USING A MINIATURE REACTION CARTRIDGE

被引:60
作者
TOTTY, NF [1 ]
WATERFIELD, MD [1 ]
HSUAN, JJ [1 ]
机构
[1] MIDDLESEX HOSP,LUDWIG INST CANC RES,RECEPTOR STUDIES GRP,LONDON W1P 8BT,ENGLAND
关键词
HPLC; MICROCARTRIDGE; SEQUENCING;
D O I
10.1002/pro.5560010914
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Greatly reduced automated protein sequencing degradation times of less than 25 min with concurrent on-line phenylthiohydantoin (PTH) derivative analysis times of less than 16 min have been achieved by using a miniaturized reaction cartridge with optimized chemical and analytical cycles. Using this method a wide range of standard and novel peptides and proteins have been sequenced with reproducibly high initial and repetitive cycle yields. In these accelerated analyses the recovery of the more labile PTH derivatives was markedly improved by using elevated pressure during cleavage steps and temperature programming throughout the Edman cycle.
引用
收藏
页码:1215 / 1224
页数:10
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