Authors
Irene Coin, Michael Beyermann, and Michael Bienert
Summary
Fmoc determination
Monitoring the resin loading during peptide synthesis gives useful information about the progress of the assembly. This can easily done by quantifying the amount of Fmoc removed at each deprotection/coupling cycle via spectrophotometry.
Kaiser Test
The Kaiser test1 is a qualitative test for the presence or absence of free primary amino groups, and it can be a useful indication about the completeness of a coupling step. The test is based on the reaction of ninhydrin with primary amines, which gives a characteristic dark blue colour. The test requires minimal amounts of analyte and is completed within a few minutes.
Microcleavage
When Fmoc removal data show anomalies, before performing a step that requires the use of particularly expensive materials, or after a critical step, and in general when assembling longer sequences, it may be useful to cleave and analyse a small amount of intermediate product.
Further details
The protocol was published on Protocol Exchange in 2007. To see the entire protocol, click on the source link.
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