蛋白质，肽中二级结构检测方案(圆二色光谱仪)

Application NoteCDDSSpectroscopy 2/2JWMVS-529 CD multivariate SSE analysis program JWMVS-529 CDmultivariate SSE analysis program The function of a protein or peptide is closely related toits structure. Since CD measurements probe amino acidstructure changes， CD is widely used for the structuralanalysis of protein and peptide molecules formulatedfor pharmaceutical use. CD spectral shapes reflect theabundance ratio of secondary structure motifs in proteinsand poly-peptides and secondary structure analysis of CDdata provides a fast and accurate indication of change inthese structures. The JWSSE-513 Protein Secondary Structure Analysisprogram uses the Classical Least Squares (CLS) method，including a reference spectra set of Yang1 and Reed2. TheYang reference spectra are extracted from the CD spectra ofproteins and are best suited for protein secondary structureanalysis. On the other hand， the Reed reference spectra areextracted from the CD spectra of peptides and are suitablefor the secondary structure analysis of peptides. The tworeference spectra are separate due to the aromatic sidechain residue bands that are observed in the CD spectra ofproteins but not peptides. The JWMVS-529 Multivariate SSE analysis program includesa library of 26 protein CD spectra (176-260 nm) which usea calibration model based on the spectra and created byJASCO.A Partial Least Squares (PLS) method with the latestmultivariate analysis method and Principle ComponentRegression (PCR) method are included. In the PLS and PCRmethods， the spectra rely on a few potential factor for whichthe concentration is determined. TO minimize the residualerror of the concentration， the abundance ratio of thesecondary structure is calculated. This significantly improvesthe analysis accuracy of the B-sheet motif， which has nostrong， specific CD marker band. This application note highlights the features of theJWSEE-513 and JWMVS-529 Secondary Structure Analysisprograms. Application Note Keywords Secondary structure analysis， multivariate， PLS method， PCR method，JWSEE-513， JWMVS-529， biochemistry Features ·PLS and PCR methods are much more precise multivariate analysis methods compared with the CLS method， whichhas been traditionally used for protein secondary structure analysis ·Verifies the calibration model using cross-validation (Figure 1) ·Edits the ratio of secondary structure and reference spectra ·Validates the analysis result using a F-test ·Verification of recalculation and calculated results (GLP/GMP compliant) ·21 CFR Part 11 compliant ·Includes CD spectra (176-260 nm) of 26 proteins with a calibration model based on those spectra Figure 1. Results of cross-validation using the PLS and PCR methods References 1. Yang， J. T.， Wu， C.S.C.， and H. M. Martinez， Methos in Enzymology (1986)，130，208-269. 2. J.Reed and T. A. Reed， Anal. Biochem， (1997)， 254，36-40. JASCO INC. 28600 Mary’s Court， Easton， MD 21601 USA JASCO INC.Mary’s Court， Easton， MD USATel：( Fax： (pplication Library： http：//www.jascoinc.com/applications Tel： ( Fax： ( pplication Library： http：//www.jascoinc.com/applicationsCopyright O JASCO Corporation The function of a protein or peptide is closely related to  its structure. Since CD measurements probe amino acid  structure changes, CD is widely used for the structural  analysis of protein and peptide molecules formulated  for pharmaceutical use. CD spectral shapes reflect the  abundance ratio of secondary structure motifs in proteins  and poly-peptides and secondary structure analysis of CD  data provides a fast and accurate indication of change in  these structures.