2002 60 12, 2115 2119 ACTA CHIMICA SINICA Vol 60, 2002 No 12, 2115 2119 SERS a a b b c c a Ξ, a a Ξ ( a 130023) ( b 650091) ( c 130021) (SERS), ( HBsAg) (,Ab2HBsAg) SERS,Ab2HBsAg (COO - ) HBsAg Ab2HBsAg,HBsAg (Trp), Trp HBsAg, HBsAg Ab2HBsAg,,, Surface2Enhanced Raman Spectroscopic Study on the Interaction of Hepatitis B Virus Surface Antigen and Its Monoclonal Antibody WANG, Lian2Ying a J I, Xiao2Hui a YUAN, Hang b MA, Lan b KONG, Xiang2Gui c LIU, Yi2Chun c YANG, Wen2Sheng a BAI, Yu2Bai Ξ, a LI, Tie2Jin a ( a College of Chemistry, Jilin University, Changchun 130023) ( b Kunming Yunda Monoclonal Antibody Technology Center, Yunnan University, Kunming 650091) ( c Institute of Optics, Fine Mechanics and Physics, Chinese Academy of Sciences, Changchun 130021) Abstract Surface2enhanced Raman scattering (SERS) has been employed to study the interaction of hepatitis B virus surface antigen ( HBsAg) with mouse monoclonal antibody against HBsAg (Ab2HBsAg) SERS is used to obtain spectra of HBsAg, Ab2HBsAg and their immune complex adsorbed on colloidal silver It is found that Ab2HBsAg is bound to silver colloids through the carboxyl groups, which are not on the antigen binding site of the Ab2HBsAg The vibrations from tryptophan residues of the HBsAg disappear after formation of the immune complex It is assumed that the tryptophan residues are among the antigen active sites of the HBsAg and play an important role in the interaction between Ab2HBsAg and HBsAg Keywords SERS, antigen, monoclonal antibody, silver nanoparticles ( ), [1 ], (SERS), 10 6 10 7, [2,3 ], [4, ] 1997 Ξ E2mail : yubai @mail jlu edu cn Received May 17, 2002 ; revised July 15, 2002 ; accepted September 1, 2002
2116 Vol 60, 2002 Nie [5] SERS, 10 14 10 15, SERS Grabbe Buck [6] SERS, ( Trp ),, Ab2HBsAg (Tyr), Dou [7 ] 2 SERS, SERS Ab2HBsAg, HBsAg Ab2HBsAg/ HBsAg SERS 1, SERS, ( HBsAg) (Ab2HBsAg), Ab2, 2 2 HBsAg (COO - ), ; HBsAg, 1670 cm - 1 (Amide I), Trp, Tyr COO - ; Ab2HBsAg HBsAg SERS,, Trp, HBsAg, SERS 1 1 1 HBsAg( 23000) Ab2HBsAg( IgG 1, 160000) HBsAg Ab2HBsAg/ HBsAg SERS 1245 cm - 1 HBsAg SERS 1542 cm - 1, Amide III Amide II [6], 18 M cm JOBIN YVON UV2LabRam Infinity, HBsAg 215 cm - 1, 488 nm 223 1 HBsAg, 20 mw, SERS 1 1594 1183 cm - 1 10 s, 20 1 2 8a 1295 809 cm - 1, Tyr SERS Lee Meisel [8 ] : 100, SERS ml 10-3 mol L - 1 AgNO 3, Tyr HBsAg, 2 ml 1 %( wt %), 1 h,tyr Grabbe Buck [6], 1 3,, 20 L 8219 g/ ml Ab2HBsAg 1 4 ml ; 200 L 2 g/ ml HBsAg 400 L ; 40 L HBsAg 40 L Ab2HBsAg, Ab2HBsAg/ Ab2HBsAg, HBsAg SERS, 1670 cm - 1 Amide I, [4,6,7],Amide II, Amide III Amide I,,, Ab2 Trp Tyr 2 1 HBsAg SERS [16],, ( Tyr),, Tyr :1389 cm - 1 COO -, 917 cm - 1 C COO -, 566 cm - 1 COO - (wagging) HBsAg 4 ml,,1389 917 cm - 1,
No 12 : SERS 2117 1 Ab2HBsAg, HBsAg Ab2HBsAg/ HBsAg SERS Table 1 Assignment of Raman bands of Ab2HBsAg, HBsAg and Ab2HBsAg/ HBsAg in silver colloids Frequency shift/ cm - 1 HBsAg Ab2HBsAg Ab2HBsAg/ HBsAg 531 w 566 w COO - wagging 13 731 w 605 w 716 w 718 mw COO - 11 760 w Tyr (ring) 6 809 ms 810 w Tyr (ring) 9 857 w Tyr (ring) 9 917 ms - C2COO 10 935 m 933 w - C2COO 10 1008 m 1009 m Phe (ring) 11 1068 w C N 1131 w 1133 w 1135 m r (CH 2 ) 12 1183 s 1179 m Tyr (ring) 6, 9, 12 1245 w 1242 m Trp 1295 w Tyr (ring) 6 1358 w Trp (ring) 6, 13 1376 s Trp (ring) 9 1389 sm 1389 s 1396 s - s, COO 12, 14 1448 w Trp 6, 12 1488 w Trp (ring) 6, 11, 12 1542 w Trp (ring) 13 1594 s 1592 w Tyr (ring) 14 1625 s 1624 br 1624 w Trp (indole ring), (H2 O) 9, 11, 15 v very ; w weak ; m medium ; s strong ; br broad stretch ; deformation ; r rocking Trp tryptophan ; Tyr tyrosine ; Phe phenylalanine 1 HBsAg SERS COO -,935 cm - 1 C Figure 1 SERS spectra of HBsAg adsorbed on silver COO -,716 cm - 1 COO -, nanoparticles Ab2HBsAg Asp, Glu C2 COO - Ab2HBsAg 1 1625, 1542, 1488, 1376, 1358 760 SERS Trp [9 ], Trp, 1 1068 cm - 1 C N, 1131 cm - 1 C CN, 3274 2980 cm - 1 NH 2 NH + 3, HBsAg 2 2 Ab2HBsAg SERS 2a Ab2HBsAg SERS,, 1389 cm - 1 COO - 1008 cm - 1 ( Phe) cm - 1 (Trp),,
2118 Vol 60, 2002 Phe 1600 cm - 1, Phe Ab2HBsAg SERS, 1624 cm - 1 Tyr, Phe Ab2HBsAg/ HBsAg SERS Tyr 1006 cm - 1[13], SERS HBsAg, Phe HBsAg SERS, 1625, 1542, 1488, 1376, 1358 760 cm - 1 Trp HBsAg Ab2HBsAg,SERS Trp HBsAg Trp [16 ], HBsAg Ab2HBsAg, HBsAg Trp Trp,, Trp Trp HBsAg, Ab2HBsAg,Trp Ab2HBsAg " ( ), 2 Ab2HBsAg (a) Ab2HBsAg/ HBsAg (b) SERS Figure 2 SERS spectra of Ab2HBsAg ( a) and Ab2HBsAg/ HBsAg (b) adsorbed on silver nanoparticles 3 SERS :HBsAg Tyr, Trp ;Ab2 HBsAg 2 3 HBsAg Ab2HBsAg SERS HBsAg Ab2HBsAg Ab2HBsAg IgG 1, HBsAg,Ab2HBsAg,, HBsAg Ab2HBsAg,, ;,HBsAg HBsAg 2b Ab2HBsAg/ HBsAg SERS Trp, Trp Ab2HBsAg HBsAg HBsAg SERS -,, 1396, 933 References 718 cm - 1 ( - s,coo ), ( C C ) ( - COO ) 1 Colloidal Gold : Principles, Methods and Applications, Vol HBsAg Ab2HBsAg, 3, Ed : Hayat, M A, Academic Press, New York, 1989 COO - - C COO 917 935 cm - 1 2 Mann, S ; Shenton, W ; Li, M ; Connolly, S ; Ab2HBsAg/ HBsAg SERS, - C COO Fitzmaurice, D Adv Mater 2000, 12, 47 933 cm - 1, Ab2HBsAg/ HBsAg 3 Niemeyer, C M ; Burger, W ; Peplies, J Angew Ab2HBsAg Chem, Int Ed 2000, 37, 2265 COO - 4 Ahern, E S ; Garrell, R L Langmuir 1991, 7, 254 2b 1624 cm - 1 5 Nie, S2M ; Emory, S R Science 1997, 275, 1102 1592 cm - 1 6 Grabbe, E S ; Buck, R P J Am Chem Soc 1989,, 111, 8362 810, 850 1179 cm - 1, 7 Dou, X ; Yamaguchi, Y ; Yamamoto, H ; Doi, S ; Tyr HBsAg SERS,Tyr Ozaki, Y J Raman Spectrosc 1998, 29, 739, 8 Lee, P C ; Meisel, D P J Phys Chem 1982, 86,
No 12 : SERS 2119 3391 9 Parker, F S Applications of Infrared, Raman, and Resonance Raman Spectroscopy in Biochemistry, Plenum Press, New York, 1983, Chapter 3 10 Suh, J S ; Moskovits, M J Am Chem Soc 1986, 108, 4711 11 Herne, T M ; Ahern, A M ; Garrell, R L J Am Chem Soc 1991, 113, 846 12 Grabbe, E S ; Buck, R P J Electroanal Chem 1991, 308, 227 13 Dollish, F R ; Fateley, W G ; Bentlek, F F Characteristic Raman Frequencies of Organic Compounds, John Wiley and Sons, New York, 1973, Chapter 9 14 Lee, H I ; Kim, M S ; Suh, S W Bull Korean Chem Soc 1988, 9, 218 15 Lee, H I ; Suh, S W ; Kim, M S J Raman Spectrosc 1988, 19, 491 16 The primary structure of Hepatitis B virus surface Antigen : MENIT SGFLG PLLVL QAGFF LLTRI LTIPQ SLDSW WTSLN FLGGT TRCLG QNSQS PTSNH SPTSC PPTCP GYRWM CLRRF IIFLF ILLLC LIFLL VLLDY QGMLP VCPLI PGSST TSTGP CRTCT TPAQG TSMYP SCCCT KPSDG NCTCI PIPSS WAFGK FLWEW ASARF SWLSL LVPFV QWFVG LSPTV WLSVI WMMWY WGPSI LSPFL PLLPI FFFLW VYI (A0205178 CHENG, B ; DONG, H Z )