ACTA CHIMICA SINICA. ( HBsAg). SERS

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1 , ACTA CHIMICA SINICA Vol 60, 2002 No 12, SERS a a b b c c a Ξ, a a Ξ ( a ) ( b ) ( c ) (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 ) ( b Kunming Yunda Monoclonal Antibody Technology Center, Yunnan University, Kunming ) ( c Institute of Optics, Fine Mechanics and Physics, Chinese Academy of Sciences, Changchun ) 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), , [2,3 ], [4, ] 1997 Ξ E2mail : jlu edu cn Received May 17, 2002 ; revised July 15, 2002 ; accepted September 1, 2002

2 2116 Vol 60, 2002 Nie [5] SERS, , 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 HBsAg( 23000) Ab2HBsAg( IgG 1, ) 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 HBsAg, 20 mw, SERS cm s, a 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,, cm - 1,

3 No 12 : SERS 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 w 605 w 716 w 718 mw COO w Tyr (ring) ms 810 w Tyr (ring) w Tyr (ring) ms - C2COO m 933 w - C2COO m 1009 m Phe (ring) w C N 1131 w 1133 w 1135 m r (CH 2 ) s 1179 m Tyr (ring) 6, 9, w 1242 m Trp 1295 w Tyr (ring) w Trp (ring) 6, s Trp (ring) sm 1389 s 1396 s - s, COO 12, w Trp 6, w Trp (ring) 6, 11, w Trp (ring) s 1592 w Tyr (ring) 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 , 1542, 1488, 1376, SERS Trp [9 ], Trp, cm - 1 C N, 1131 cm - 1 C CN, cm - 1 NH 2 NH + 3, HBsAg 2 2 Ab2HBsAg SERS 2a Ab2HBsAg SERS,, 1389 cm - 1 COO cm - 1 ( Phe) cm - 1 (Trp),,

4 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, 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 cm Mann, S ; Shenton, W ; Li, M ; Connolly, S ; Ab2HBsAg/ HBsAg SERS, - C COO Fitzmaurice, D Adv Mater 2000, 12, 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 Nie, S2M ; Emory, S R Science 1997, 275, cm Grabbe, E S ; Buck, R P J Am Chem Soc 1989,, 111, , 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,

5 No 12 : SERS 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, Herne, T M ; Ahern, A M ; Garrell, R L J Am Chem Soc 1991, 113, Grabbe, E S ; Buck, R P J Electroanal Chem 1991, 308, 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, Lee, H I ; Suh, S W ; Kim, M S J Raman Spectrosc 1988, 19, 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 (A CHENG, B ; DONG, H Z )