利用HTCD系统测量CD光谱

自动测量系统由于能够自动分析多个样本，节省时间和人力，并消除任何可能的人为错误，已引起制药公司越来越多的关注。由于CD光谱学可以让用户轻松地获得有关蛋白质和多肽的结构信息，因此自动化测量系统可以用于优化大量样品的筛选过程。该应用说明表明，三种不同的光谱学技术（CD、吸收和激发）可以在几分钟内测量出高质量的数据。J-1500与HTCD系统相结合，允许超过100个样本被自动测量，而无需人工操作，节省了时间，并消除了人为误差。Application NoteCDSSpectroscopy CD spectrum measurement usinghigh-throughput CD2/3Application Note CD spectrum measurement using high-throughput CD Introduction Au t o m atic me a su r eme n t systems have been dra win g in cr e asin g a t t e nt i on a m ong ph ar m a c e u ti c als c o mp a ni e s du e t o t h ei r a bilit y t o an a ly se multiple sample s a u t o m ati c a l ly，saving b o th ti m e a n d m a n p owe r and el i m in ati ng any p o s s ib le h u m an er r or . Si n c e CD spe ctr osco p y al lo w s th e u s er t o e a s i l y o btain stru c tu r a l in f o rmati o n ab o u t pr ot e i n s an d pe p tides ， au t oma t ed meas u rement sys t em s can be u sed t o o ptimi z e the scr e e n i n g p roc e s s of a l a r g e n umb e r of s ampl es . JASCO J-1500 high-throughput CD system View product information at w ww.j a sc oin c.c o m Th is applic a ti o n n ote s i n t r od u c es th e Hig h -Th r o u g hp ut C D syste m fo r a u tomatic C D meas u rements of m u ltiple pr otein s a m pl es . Keywords J -1500， cir c u l a r d i c h r o i s m ， A u t osa mpl er， ASU -800，F M O -427， Emission monoc h r o mator ， excitation ， emission ， Hi g h -Th rou ghp u t ， HTCD， pr oteins， bi oc h em i st ry ， ph a rm a ce u tic a ls Experimental Data acquisition interval 0.1nm Response time 2 seconds Spectral bandwidth 1nm Scan speed 100 nm/min Accumulations 4 times Path length 1mm Emission wavelength 350 nm Sensitivity 850V Results First the sequence o f opera ti on a n d data acq u isit i on m u s t b e de f ined i n advance. The max i m u m nu mb er of sam pl es tha t ca n b e a n a ly z e d a u t o m ati ca lly i s 192. Th e s y s t e m w il l t h e n c a rr y o u t CD measu r e m e n ts au tom a t icall y a n d sav e th e d ata s e r i e s. The fol l ow in g sequ e n c e is au t om ati ca ll y per f orm e d by th e HT CD s y s t em ： 1. Load in g of m easuremen t condition 2. B a seline co rr ection 3. S a mp l e 1 mea s u remen t (c y t o c h r ome c ) 4. Wash in g 5. S a mple 2 measu r e m e n t (lysozyme) 6. W ashin g 7. Sample 3 measu r e m e n t (h u ma n ser u m a lb u min ) 9. Sa mp l e 4 m easu r e m en t (hemo g l o bin) 10. Wash i n g Figure 1. Display of sequence setting The JF LC-499 CD/emis s i o n flo w cell was u sed to obtain bo th CD a n d exci t a t io n spec tr a. Th e r esu lt s o f the au t omatic me a su r em e n t s of CD ， ab sorb a n c e ， a n d excit a ti o n s p e ctr a f or f o ur 0.1 mg/mL pr o tein s ampl es ar e sh o w n b e lo w . Th e se high spee d measu r e m e n ts t oo k a t ot a l o f 3 min u te s p er sampl e (ea c h s c an i s 45 seco n ds an d t h ere are 4 a c c u m u lati o n s). JASCO INC. 28600 Mary's Cour t ， E asto n ， MD 21601US A Figure 2. CD (top)， absorbance (middle)， and excitation (bottom) spectra of cytochrome c. Figure 4. CD (top)， absorbance (middle)， and excitation (bottom) spectra of human serum albumin. Figure 3. CD (top)， absorbance (middle)， and excitation (bottom) spectra of lysozyme. Figure 5. CD (top)， absorbance (middle)， and excitation (bottom) spectra of hemoglobin. Conclusion Th is ap p l i c a ti o n n ot e de m o n strates th at h igh qu a lity da t a ca n b e me a s u re d fo r thr e e d i ffe r e n t s pe ctr o sc o p y tec h n iqu e s (CD， abso rp tio n ， a n d e x c i t a ti o n) in a mat t er of m i n u t es . T h e J-1500 coupl ed w i th th e HTC D s yst e m allows over 100samples to be measu r ed a u tomatically w itho u t hu m a n o p e r atio n ， saving t i me and elimi n ating h u ma n error. JASCO INC. 28600 Mary's Cour t ， E asto n ， MD 21601US A T el ： (800) 333-5272， Fa x ： (410) 822-7526 Appli c a ti o n Lib r ary： htt p：//www.jas co in c .c o m/a pp licatio n s