Immunosensors

Description M t
Immunosensors hnh ng trn nguyn tc rng cc phn ng min dch ca cc loi sinh hc nht nh (thng l vi khun) vi cht gy nhim s to khng th, do c th o c. gim chi ph v thi gian cn thit phn tch chnh xc cc mu lnh vc nc v t b nhim vi cc hp cht n, chng hn nh trinitrotoluene (TNT) v Royal Demolition n (RDX) , hai immunosensors c pht trin. H l nhng si quang cm bin sinh hc v min dch lin tc dng chy cho trn nhng ch kim tra v gim st cc cht gy nhim. C hai b cm bin xc nh mc nhim bng cch o mc hunh quang hot ng gy ra bi s ra i ca mt mu sinh hc vo h thng. Cc si quang cm bin sinh hc phn t cht gy nhim hot ng khi cnh tranh vi cc khng th hunh quang trn cm bin. S suy gim trong hot ng hunh quang gy ra bi cc cht nhim vo cc ch lin kt khng th tng ng vi mc nhim. Cc quy trnh lin tc min dch hot ng khi cc phn t cht gy nhim di di cc khng th hunh quang c t trn mt s h tr vng chc. Khi khng th di di c pht hin, chng tng ng vi t l thun vi mc nhim.

Hn ch v mi quan tm
Phng php s dng cc cm bin sinh hc da trn cc k thut xt nghim min dch hunh quang pht hin thuc n TNT v RDX c mt s lng ln cc tch cc cc gi tr sai cho cc kho nghim TNT. Khng thng tin c trnh by v cc cng ngh ny gii quyt nhng hn ch v quan tm y . Khng r liu cc cm bin ny c hiu qu trong cc hp cht pht hin khc vi RDX v TNT Trong khi cc cm bin c th c p dng cho cc kim tra ha cht, n l khng r rng nh th no h s c s dng theo di. C v nh cc cht gy nhim phi tip xc trc tip vi cm bin. Xem

thm thng tin cn thit v cch cc cm bin c th c sp xp phc v cho mt chc nng gim st.

ng dng
Cc cm bin l cc cng c phn tch pht hin thuc n TNT v RDX trong t v nc ngm . C hai b cm bin c cm tay, nhanh chng (2-10 pht mi phn tch), v nhy cm (cc b phn thp mi ln pht hin nng t USD).

Tnh trng pht trin cng ngh

Cc cm bin sinh hc si quang v lin tc min dch dng chy, pht trin Phng th nghim Nghin cu Hi qun (NRL) phi hp vi cc nghin cu quc t, s c s dng ti mt trang web pht hin cht n TNT v RDX. Cc cm bin c coi l d n th im.

Web Links Lin kt web

http://www.serdp-estcp.org/content/download/3607/58263/file/CU199713-CP.pdf http://www.serdpestcp.org/content/download/3607/58263/file/CU-199713-CP.pdf

Ti nguyn khc v cuc biu tnh

lnh vc th nghim rng ri (ba i vi nc ngm v mt cho t) c tin hnh ti mt s trang web. Cc trang web nc ngm, cc tu ngm hi qun Bangor c s (Washington), Ha cht Umatilla Depot (Oregon), v cc trc hi qun Surface Warfare Center (Indiana), ang trn EPA Hoa K Danh sch u tin quc gia ("Superfund"). Nhn chung, kt qu cho cc cm bin sinh hc cho rng cc cng c cng ngh y ha hn lnh vc s yu cu pht trin thm trc khi chng c s dng thch hp cho trng.

Immunosensors

Cm bin sinh hc c th c s dng kt hp vi cc enzyme lin kt xt nghim min dch (ELISA). . Cc nguyn tc ng sau k thut ELISA c th hin trong hnh 6,9 . ELISA c s dng pht hin v khuych i mt phn ng khng nguynkhng th, s lng khng nguyn lin kt enzym lin kt vi cc khng th c nh c xc nh bi s tp trung tng i ca cc v lin hp khng nguyn min ph v nh lng bng t l ca phn ng enzym Enzym vi s doanh thu cao c s dng t c phn ng nhanh. nhy ca xt nghim ny c th c tng cng hn na bng cch s dng xc tc cc phn ng enzyme, trong cung cp cho phn ng ln hn v bn cht, v d, nhng ngi tng cho n, hunh quang mu cao hoc cc sn phm bioluminescent Kho nghim b dng c s dng k thut ny ang c sn cho mt phm vi rng ln ca cc phn tch.

. Hnh 6,9. Nguyn tc ca mt cnh tranh trc tip ELISA. (I) Khng th, c th i vi cc khng nguyn ca li sut l c nh trn b mt ca ng. Mt hn hp ca mt khon tin gi ca-enzyme lin hp cng vi nng khng nguyn l ca khng nguyn mu c t trong ng v c cn bng. (Ii) Sau mt thi gian ph hp vi cc khng nguyn v khng nguyn enzyme lin hp s c phn phi gia cc quc gia b rng buc v min ph ph thuc vo nng tng i ca h. ( (Iii) Cha cam kt vt liu c ra ra v b i. Lng-enzyme lin hp khng nguyn l rng buc c th c xc nh bng t l ca cc phn ng enzym tip theo. Gn y, cc k thut ELISA c kt hp vi cm bin sinh hc, hnh thnh immunosensors, tng phm vi tc , v nhy cm. Mt cu hnh min dch n gin c th hin trong hnh 6,10 (a) , ni m b cm bin s ch n thun l thay

th vic pht hin h thng o mu truyn thng. Tuy nhin tin tin immunosensors hn ang c pht trin ( Hnh 6,10 (b) ) m da vo vic pht hin trc tip ca khng nguyn gn vi b mt ph khng th ca cm bin sinh hc ny. p in v cm bin sinh hc da trn FET c bit ph hp vi cc ng dng nh vy.

Hnh 6,10. Nguyn tc ca immunosensors. (a (A) (i) Mt ng c ph mt lp (c nh) khng nguyn. Phn vt mc khng th-enzym lin hp c th c t trong ng v c cho php rng buc. (A) (ii) Sau mt thi gian ph hp vi bt k ti liu khng lin kt c ra i.. (A) (iii) Cc gii php khng nguyn analyte l thng qua thnh ng, rng buc v gii phng mt s cc-enzyme lin hp khng th ph thuc vo nng khng nguyn ca cc. Lng-enzyme lin hp khng th pht hnh c xc nh bi cc phn hi t cc cm bin sinh hc. (B) (i) b chuyn i A l bao vi (c nh) khng th, c th i vi cc khng nguyn ca tm. u d c ngm trong dung dch c cha mt hn hp ca mt khon tin gi ca-enzyme lin hp khng nguyn cng vi nng khng nguyn l ca mu. (B) (ii) Sau mt thi gian ph hp vi cc khng nguyn v khng nguyn enzyme lin hp s c phn phi gia cc quc gia b rng buc v min ph ph thuc vo nng tng i ca h. (B) (iii) Cha cam kt vt liu c ra ra v b i.. Lng-enzyme lin hp khng nguyn rng buc c xc nh trc tip t tn hiu ti np.

Immunosensors Cm bin sinh hc c th c s dng kt hp vi enzyme - lin kt xt nghim min dch (ELISA). ELISA c s dng pht hin v khuych i mt phn ng khng nguyn-khng th, lng enzyme - khng nguyn lin kt vi cc khng th c nh c xc nh bi s tp trung tng i ca cc khng nguyn t do v lin hp v nh lng bng t l ca phn ng enzym Enzym vi s doanh thu cao c s dng t c phn ng nhanh. nhy ca xt nghim ny c th c tng cng hn na bng cch s dng cc phn ng enzyme xc tc cho phn ng m thc cht hn, v d, nhng ngi tng cho n hunh quang, mu sc hoc cc sn phm cao bioluminescent.

Kho nghim b dng c s dng k thut ny ang c sn cho mt phm vi ng ca phn tch. Gn y, cc k thut ELISA c kt hp vi cm bin sinh hc, hnh thnh immunosensors, tng phm vi tc , v nhy cm. Trong min dch cc b cm bin s ch thay th cc h thng pht hin truyn thng o mu.. Tuy nhin nhiu hn immunosensors tin tin ang c pht trin m da vo vic pht hin trc tip ca khng nguyn gn kt vi b mt ph khng th ca cm bin sinh hc ny. N khng th sn xut mt lot cc khng th (gi l khng th n dng) c phn ng c th khng nguyn ca mt c nhn. Nu cc cht o c t khng nguyn, sau l mt khng th c th chng li n c th c sn xut v c th c kt hp vo cc h thng cm bin sinh hc. Cc cm bin sinh hc theo cch ny c th c a ra rt c th trong hnh ng ca mnh.

Immunochemical cm bin sinh hc s dng min dch c hiu vi pht hin quang ph. Hai th tc chnh c th c s dng, phng php cnh tranh bnh sandwich v phng php ny. Trong phng php u tin, khng th (c phn ng c th vi cc cht cn o, cc analyte) l c nh trn b mt ca mt microplate vi analyte ny. Sau khi ra, mt gii php c cha cc khng th lin kt vi enzyme mt c t vo cc microplate c vi cc khng th c nh analyte phc tp t nhng bc u tin. Cc pht hin lin quan n vic o quang ph mu mt sn phm ca phn ng enzym phn nh s lng cc phn t analyte v ca cc khng nguyn kt hp vi enzyme khi thm vo cc khng th c nh. Sau khi ra, cc phn ng enzyme c s dng o lng s lng cc khng th m khng c phn tch.

Vn Regenmortel et al (1998) m t cc php o khng nguyn - khng th tng tc vi cc cm bin sinh hc. Trong k thut ny, mt trong nhng i tc tng tc l bt ng trn mt chip cm bin v cc rng buc ca ngi khc c theo sau bi s gia tng ch s khc x gy ra bi khi lng ca cc loi b rng buc.. Nhng cm bin sinh hc c th c s dng lp bn chc nng (a) ca epitope v pratopes ca t bin, (b) phn tch cc thng s nhit ng lc hc ca s tng tc, (c) o nng ca sinh hc phn t ang hot ng v (d) la chn ca tu thm d chn on. Mt h thng cm bin quang hc c pht trin, c c bit thch hp cho chn on y t point-of-chm sc (Schult et al, 1999 H thng cho php cho tt c cc loi nh dng kho nghim immunochemical v bao gm chip cm bin dng mt ln v mt thit b c quang hc-out. H thng ny cho php xc nh chorionic gonadotropin hormone thai (hCG), trong huyt thanh ca con ngi vi mt gii hn pht hin ca 1 ng / ml. H thng ny cng c tm thy l hu ch cho vic kho nghim ca theophylline trong huyt thanh bng cch s dng mt loi xt nghim min dch cnh tranh.

AC voltammatric immunosensors dn carbon da trn enzyme c pht trin bi al FernandezSanchez v cng s (2000) xc nh IgG ca con ngi. Th ng hp th ca thuc th immunochemical thch hp c thc hin trn b mt in cc. Phosphatase kim - nhn globulin min dch l tracer c s dng trong cng vic ny, 3 - indoxyl phosphate l mt cht enzyme rt ph hp pht hin in ca cc phn ng mi quan h tng ng. Cc thy phn ca phn t ny to ra dimer chm. Sn phm ny c pht hin bng cch

xen voltammetry hin li dng vic hp th v vn c tnh electrodic rng cuc trin lmPhng php ny c p dng cho cc thit k ca hai min dch khc nhau cho vic xc nh IgG ca con ngi. Benkert et al. ( (2000) pht trin mt phng php ELISA creatinine v da trn mt khng th amperometric cm bin creatinine vi mt gii hn pht hin trong khong nanomolar. Creatinine khng th c th c to ra v c s dng xc nh creatinine c nhy cao v c th immunochemical. Creatinine l derivatized ti N3 v kt KLH protein vn chuyn. Trn c s immunogen ny, khng th n dng c pht trin bi cng ngh hybridoma xc nh creatinin, cc mu creatining c vi B90-AH5 v chng IgG (chut) - lin hp glucose oxidase v p dng cho cc t bo o. Sau mt bc ra, ng c b sung v hydrogen peroxide c sn xut c ng k ti Eappl = V m 600 vs Ag / AgCl. Da trn cm bin sinh hc xt nghim sng lc immunochemical pht hin cc sulphadiazine (SDZ) v sulphamethazine (SMT) trong c chit xut t ln c pht trin (Bjurling et al, 2000.). iu ny chun b n gin v d hiu cho php ln n 40 mu c x l v phn tch trong mt ngy.. Cc gii hn pht hin cho cc xt nghim c tm thy l 5,6 ng g-1 cho SDZ v 7,5 ng g-1 cho SMT. Enzym r rng s c s dng rng ri hn trong tng lai v iu ny s c phn nh trong s cc enzyme c sn trn mt quy m (v nghin cu) cng nghip, a dng ca cc phn ng xc tc v phm vi ca iu kin mi trng, theo h s hot ng. enzyme thnh lp s c a vo s dng mi v cc enzyme mi c pht hin trong hc sinh hc ca h hoc c sn xut theo thit k s dng cng ngh enzyme s c s dng xc tc cc phn ng cha c khai thc.. y ch l s bt u ca thi i cng ngh enzym.

Immunosensors c da trn cng nhn c th v nhy cm cao ca cht phn tch (antigens) with antibodies (see picture below). (Khng nguyn) vi cc khng th (xem hnh bn di). ch yu sn xut trong cc phn ng min dch th cp, hoc cc mnh ca IgG l predominanteley c s dng trong cc ng dng min dch. Tuy nhin, globulin min dch khc cng c sn.