Peptides useful as internal standards for microsequencing...

G - Physics – 01 – N

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G01N 33/68 (2006.01) C07K 1/02 (2006.01) C07K 1/12 (2006.01) C07K 2/00 (2006.01) C07K 14/00 (2006.01) C12Q 1/37 (2006.01)

Patent

CA 2140891

The present invention provides a novel internal standard for amino acid sequencing which contains a peptide consisting of unnatural amino acid res- idues, such as ornithine, norvaline, norleucine and .alpha.-aminobutyric acid, that is capable of being sequenced si- multaneously with an unknown pep- tide or protein without interfering with the analysis. The internal standard peptide has an amino acid sequence containing at least two different unnat- ural amino acid residues having reten- tion times distinct from the corre- sponding retention times for natural amino acid residues. Information de- rived from the sequencing of the inter- nal standard allows determination of repetitive yield, lag, N-terminal block- age and discrimination between blank cycles caused by missed injection and blank cycles caused by faulty delivery of chemicals during the sequencer reactions. The present invention further provides novel synthetic control peptides containing from about 3 to 100 natural amino acid residues that are designed for use in monitoring the proper operation of amino acid se- quencers and to monitor peptide or protein cleavage reactions. The control peptide, or mixture of control peptides, are designed to obtain data for many or all common, uncommon and difficult to measure amino acids within 15 sequencer cycles and to pro- vide cleavage sites for at least 4 different amino acide cleavage reactants.

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