A ( SPA ) Su lfo succin im idyl 62[ 3 2( 22pyridyldith io ) p rop ionam ido ]hexanoate (Su lfo2l C2SPD P) C E2m ail: w
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1 V o l. 25 N o CH EM ICAL JOU RNAL O F CH IN ESE UN IV ERS IT IES C 1, 1, 1, 1, 2, 3 (1., ; 2., ; 3. 26, ) A T 10M H z, C, 2 5 2C m IU gl C nggml, C., (r ).,,,,,,. ; ; ; C R ; T P A (2004) Β,, Β C [1 ]., ; C,, C.,, Β, C., C [2 ]., C.,,,, [3 ].,,,, [4, 5 ]. [6, 7 ], 2C. A ( SPA ) Su lfo succin im idyl 62[ 3 2( 22pyridyldith io ) p rop ionam ido ]hexanoate (Su lfo2l C2SPD P) C, 2C,.., A T 10 M H z, 1310 mm 1310 mm : : ( : 962A ) ( : ) ( : 98M 064, 01M A 180, 01LO 065). : (1955 ),,,,. E2m ail: w eilingfu@yahoo. com
2 N o. 1 : 2C 45, 410 mm, ( 26 ). ( ), 610 mm,, 10 mm. 2 5 ( 26 ), T TL ;, 2 5 PESA D erecto r23000 ( ). PESA PCL 2836 ( ) Pg ( ). ( 516 m ggml, PBS ph = 710) C ( 310 m ggml, PBS ph = 710) B iodesign ; Su lfo succin im idyl 62[ 3 2( 22 pyridyldith io)p rop ionam ido ]hexanoate (Su lfo2l C2SPD P ) p ro tein A (SPA ) P ierce ; C ( ),. A. R [ 8 ]. N [ 9 ],, 3 ΛL Su lfo2 L C2SPD P, 60 m in, 25 mmo lgl (D T T ) 2 ΛL, 30 m in.,,. 1% BSA 5 ΛL, 0115 mo lgl PBS (ph = 714) 3 5. N 2, SPA 2 m ggml SPA (PBS, ph = 710) 5 ΛL, PBS 3. 3 ΛL SPA, 30 2 h, 1% BSA 5 ΛL, PBS 3, N 2, , 2, 610 mm.,,,,,, ; , F ig. 1 Schematics illustrating the con struction of 2 5 model of quartz crystal m icro-array, 1., C, , 1% BSA. 100 ΛL (PBS ph = 712), 37, 2 5 f 1; 10 ΛL C,, 40 m in, f 2, f = f 1- f 2. 3.
3 4 6 V o l , 810 mo lgl 80 ΛL, 5 m in, ph = 710 PBS 5, SPSS x 2 t f ( f = f 0- f 1, f 0 ), 1.,, Su lfo2l C2SPD P SPA, ; ( 2 ), [ 8 ]. C m ggml,, Su lfo2l C2SPD P. F ig I2 125 I2C 30 ΛL 2 Relation sh ip between concen tration s of an tiin sulin an tibody and frequency sh if ts of quartz crystals, 4, PBS (ph = 710) 3., cpm ( 1).,, 125 I2 125 I2C ; cpm,, C SPA (P < 0101),. Table 1 Effects of two methods on the amoun t and the coherence of an tibody immobilization (x s) M ethods F requency sh ift a gh z CV (% ) Radioactive count (cpm ) CV (% ) IA b CA c IA b CA c IA b CA c IA b CA c T h io l m ethod SPA m ethod a. Caused by antibody immobilization; standard deviations of 10 results fo r each m ethod, P < 0101 vṡ th io l m ethod and SPA m ethod; b. Insulin antibody (516 m ggml ) ; c. C pep tide antibody (310 m ggml ) (T SH ) (FSH ) (L H ), 2., SPA. Table 2 Nonrespon se of p iezoelectr ic in sulin-c peptide m icro-array immunosen sor to other an tigen s in human serum (x s) 3 A ntigens TSH FSH LH f (T h io l m ethod) gh z f (SPA m ethod) gh z f : F requency sh ifts caused by T SH (100 ΛIU gml ), FSH (200 m IU gml ) and L H (250 m IU gml ). Standard deviations of 5 results fo r each m ethod, P < 0105 vṡ th io l m ethod and SPA m ethod.. SPA. [10 ], SPA,. SPA,
4 N o. 1 : 2C 47. Su lfo2l C2SPD P SPA, ( SH ), A u S,,,,,, LB., Su lfo2l C2SPD P,,,, SPA [11 ]. SPA A u SPA 2,. SPA, IgG F c,, F ab,,, [10 ]., Su lfo2l C2SPD P SPA,,,., Su lfo2l C2SPD P, C [ 8 ]. 37, 2.,. 2,, 2,, C, 37, 2 ( 3 4)., 2 40 m in,,. 2,,,., 40 m in 2C, EL ISA 2. F ig. 3 Tendency of frequency change with time of in sulin p iezoelectr ic immunosen sor Insulin concentration: 10 m IU gml. F ig. 4 Tendency of the frequency change with time of C peptide p iezoelectr ic immunosen sor C pep tide concentration: 1125 nggml ph,. [12 ], 50%,., ph PBS, 10,, C ( f = f 1- f 2; f 1 ) 3,. C, ( f ), ( 5 6)., m IU gl, C nggml, C
5 4 8 V o l. 25, 215 m IU gl C nggml,., 215 m IU gl nggml C., : f = lnx (P < 0101) ; C f = lnx (P < 0101). F ig. 5 Normalized respon se curve of p iezoelectr ic m icro-array imm unosen sor to in sul in concen tration s 2. 4 F ig. 6 Normalized respon se curve of p iezoelectr ic m icro-array imm unosen sor to C peptide concen tration s 2C 63.,,, , (P > 0105) mo lgl, C,,. 5, 70%,.,, 5., Su lfo2l C2SPD P SPA,,,,,.., 5,. 2C 63,. 2C,,. [ 1 ] V an N. J.. L ancet[j ], 2001, 357 (9 273) : [ 2 ] W righ t P. R., M ills J., Choo K. E. et a l.. W est Indian M ed. J. [J ], 2000, 49 (2) : [ 3 ] Stick le D. F., M cn eil S. E.. C lin. Ch im. A cta[j ], 1999, 1283 (1 2) : [ 4 ] CH EN Ze2Zhong ( ), WAN G Ke2M in ( ), CH EN Yong2Kang ( ) et al.. Chem. J. Ch inese U niversities( ) [J ], 2002, 23 (6) : [ 5 ] WU Zhao2Yang ( ), H E Chun2P ing ( ), WAN G Sh i2p ing ( ) et al.. Chem. J. Ch inese U niversities( ) [J ], 2003, 22 (4) : [ 6 ] T akash i A., M asayo sh i E.. Senso rs and A ctuato rs B [J ], 2000, 82: [ 7 ] WAN G L ei2l ei ( ), FAN G H ui( ), ZEN G Q ing2yin ( ) et al.. J. A nal. Sci. ( ) [ J ], 2001, 17 (5) : [ 8 ] ZHAN G Bo ( ), FU W ei2l ing ( ), J IAN G T ian2l un ( ) et al.. A cta A cadem iaem edicinaem ilitaris T ertiae ( ) [J ], 2002, 24 (1) : [ 9 ] Park İ S., Kim N.. B io senṡ B ioelectron. [J ], 1998, 13: [ 10 ] PE I Ren2Jun ( ), HU J i2m ing ( ), HU Yi( ) et al.. Chem. J. Ch inese U niversities( ) [J ], 1998, 19 (3) : [ 11 ] N irm alya K., Chak i K.. V ijayamohanan et a l.. B io senṡ B ioelectron. [J ], 2002, 17 (1 2) : 1 12
6 N o. 1 : 2C 49 [ 12 ] ZHAN G Bo ( ), FU W ei2l ing ( ), J IAN G T ian2l un ( ) et al.. A cta A cadem iaem edicinaem ilitaris T ertiae ( ) [J ], 2002, 24 (1) : 8 11 Stud ies on P iezoelectr ic In sulin-c Peptide M icro-array Imm unosen sor ZHAN G Bo 1, FU W ei2l ing 13, M AO Q iong2guo 1, ZHAN G Xue 1, TAN G W an2l i 2, YU Fan 3 (1. D ep artm ent of L aboratory, S ou thw est H osp ital, T h ird M ilitary M ed ical U niversity, Chong qing , Ch ina; 2. D ep artm ent of B ioeng ineering, Ch ina J ialing G roup, Chong qing , Ch ina; 3. T he 26th R esearch Institu te of E lectronics, M inistry of Inf orm ation Ind ustry, Chong qing , Ch ina) Abstract A new 2 5 type of p iezoelectric in su lin2c pep tide m icro2array imm uno sen so r w as developed fo r the determ ination of in su lin and C pep tide. T he energy converters w ere 10M H z A T 2cu t quartz crystals w ith go ld2coated electrodes. T he monoclonal an tibodies of in su lin and C pep tide w ere imm ob ilized on to the su rfaces of crysta ls go ld elect rodes resp ect ively, then p iezoelect ric in su lin2c pep tide m icro2array imm uno sen so r w as con structed by 2 5 type of immob ilization clamp ing apparatu s. T he effects of m ethods, w o rk concen tration s, the amoun t and coherence of immob ilization of an tibody and the respon se param eters of the m icro2array imm uno sen so r such as temperatu re, tim e2 co st and specificity etc. w ere investigated. U nder the op tim ized condition s, the experim en tal resu lts show ed p iezoelectric imm uno sen so r had good respon se to in su lin and C pep tide w ho se frequency sh ifts w ere linearly dependen t on in su lin concen tration in the range of m IU gl and C pep tide concen tration in the range of nggml. T he p iezoelectric m icro2array imm uno sen so r w ere u sed to detect in su lin and C pep tide in serum, the analytical resu lts given by th is app roach w ere in satisfacto ry agreem en t w ith tho se given by radiom etric imm unoassay (P > 0105), w ith co rrelation coefficien t s 0192 and T he p iezoelect ric m icro 2a rray imm uno sen so r fo r the determ ina t ion of in su lin and C pep tide is of h igh sen sitivity, h igh specificity, low den sity m icro2array structu re, h igh analysis speed, unnecessary labelling, simp le operation, real2tim e detection and repeated u se, etc.. It is su itab le fo r detecting in su lin and C pep tide of serum in clin ical labo rato ry. Keywords P iezoelectric imm uno sen so r; M icro2array; In su lin; C pep tide (Ed. : K, G)
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14 2 V o l114,n o12 1998 4 Ch inese Jou rnal of B iochem istry and M o lecu lar B io logy A p ṙ 1998 3 - (, 100044),,,, 125 g, :,,, Quan tif ica tion of Polym era se Cha in Reaction Products Using Solid
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2006, 26 (1): 87 92 Journal of T riticeae C rop s Ξ,,,,, ( g, 225009) : 6,,,,,,, : ; ; ; ; : S 512. 1; S 318 : A : 100921041 (2006) 0120087206 Stem Character istics of W heat w ith Stem L odging and Effects
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