Fluorometric method for quantitative determination of free amine groups in peptide-containing merrifield resins
✍ Scribed by John Garden II; Andrew M. Tometsko
- Publisher
- Elsevier Science
- Year
- 1972
- Tongue
- English
- Weight
- 418 KB
- Volume
- 46
- Category
- Article
- ISSN
- 0003-2697
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✦ Synopsis
Recent research in this laboratory has resulted in the construction of an automatic chemical reaction system (ACRS) (1) for carrying out the physical operations involved in the solid-phase chemical synthesis and/or solid-phase Edman degradation (2) of polypeptides and proteins. The ACRS reads and translates a computer-generated control tape which contains the binary code for each operation. As discussed previously, the computer could readily select reaction conditions (e.g., reaction times, deblocking times, etc.) that would be tailored for each individual amino acid in essentially the same time taken to generate a tape where all amino acids are subjected to identical reaction conditions. In order to utilize the programming flexibility of our computer methods for preparing control tapes, additional data are needed concerning the rates of incorporation of amino acids into resin-bound peptides (disappearance of free amines) and rates of deblocking resin-bound peptides (appearance of free amines). The coupling rates of amino acids have been found to vary with the identity of the incoming amino acid by Esko et al. ( 3), with the identity of the activating group by Karlsson et al. (4), and with chain length by Hagenmaier (5). Similarly, the efficiency and rate of the deblocking reaction have been found to vary with the identity of the blocking group by Ragnarsson, et al. (6) and with deblocking conditions by Karlsson et al. (7) and by Chou et al. (8).
As a result of our search for a method for determining the quantity of free amine present in a peptide-containing resin, the reported highly sensitive fluorometric procedure was developed. This method compares favorably with methods developed in other laboratories (3,9-12) in sensitivity, accuracy, and reproducibility.