Ipulatifomu Yokuthuthukiswa kwe-Aptamer

Ama-aptamer ayi-oligonucleotide enezintambo ezilodwa (i-DNA, i-RNA noma i-XNA) enesakhiwo sokunamathelana okuphezulu kanye nokucaciswa okuphezulu okubopha ngqo kuma-molecule aqondiwe njengama-antibodies, futhi asetshenziswa kakhulu ekuthuthukiseni ama-biosensors, ukuxilongwa kanye nokwelapha.

Ipulatifomu ye-aptamer enikezwa yi-Alpha Lifetech ihlanganisa izigaba ezimbili: ipulatifomu yokwenziwa kwe-aptamer, ehilela kakhulu insizakalo yokwenziwa komtapo wezincwadi we-SELEX aptamer kanye nensizakalo yokuthuthukiswa kwe-aptamer (i-DNA, i-RNA noma i-XNA), kanye nepulatifomu yokuhlolwa kwe-aptamer okuhlanganisa nezinsizakalo zokuhlola ezisekelwe kubuchwepheshe be-SELEX bamaprotheni, ama-peptide, amaseli, ama-molecule amancane, ama-ion ensimbi kanye namanye ama-molecule aqondiwe, kanye nezinsizakalo zokuhlaziya nokwenza ngcono i-aptamer.

Ipulatifomu Yokwenziwa kwe-Aptamer

Insizakalo yokuhlanganisa umtapo wezincwadi we-SELEX aptamer

Insizakalo yokwenziwa komtapo wolwazi we-SELEX aptamer ihilela kakhulu ekwakheni umtapo wolwazi oqukethe inani elikhulu lezinhlelo ze-oligonucleotide ezingahleliwe ezinezintambo ezilodwa ngokwenziwa kwamakhemikhali e-in vitro ngokwezinhlayiya eziqondiwe. Ukwakhiwa komtapo wolwazi kuyindawo yokuqala yobuchwepheshe be-SELEX, obuhlinzeka ngezinhlelo eziningi zenqubo yokuhlola elandelayo ngokwakha imitapo yolwazi emikhulu engahleliwe futhi kwandisa amathuba okuhlola ama-aptamer ahambisana kakhulu.

Ukuhlanganiswa komtapo wolwazi kuhlukaniswe kakhulu ngezinyathelo ezilandelayo:

Izinyathelo	Imininingwane Yobuchwepheshe
Thola Ama-molecule Aqondiwe	Thola ama-molecule aqondiwe okudingeka ahlolwe ama-aptamers, okungaba amaprotheni, ama-nucleic acid, ama-molecule amancane, ama-ion ensimbi, njll.
Umklamo Wokulandelana Okungahleliwe	Ubude bokulandelana okungahleliwe, ukwakheka kwesisekelo kanye neminye imingcele yaklanywa ngokwezici zama-molecule aqondiwe kanye nezidingo zokuhlola. Ngokuvamile, ukulandelana okungahleliwe kuphakathi kwamashumi namakhulu ezisekelo ngobude.
Ukuhlanganiswa Kwezilandelano Ezingaguquki	Izingcezu ze-oligonucleotide ezinezilandelaniso ezihleliwe (njengezilandelaniso ze-PCR primer) kuzo zombili iziphetho zaklanywa futhi zahlanganiswa, ezizosetshenziswa enkambisweni elandelayo yokukhulisa nokuhlola.

Umtapo wolwazi ohlanganisiwe usadinga ukucutshungulwa kabanzi ukuze kulawulwe ikhwalithi. Ukuhlushwa komtapo wolwazi kwanqunywa ukuqinisekisa ukuthi uyasebenza enkambisweni yokuhlola elandelayo. Ukuhlukahluka kanye nokunemba kokulandelana okungahleliwe emtapweni wolwazi kwaqinisekiswa ngokulandelana kanye nezinye izindlela zokuqinisekisa ukuthi ikhwalithi yomtapo wolwazi iyahlangabezana nezidingo zokuhlola.

Ngezinyathelo ezingenhla, umtapo wolwazi we-SELEX aptamer osezingeni eliphezulu futhi ohlukahlukene kakhulu ungakhiwa, onganikeza izinhla eziningi zenqubo yokuhlola elandelayo.

Izinsizakalo zokuthuthukiswa kwe-Aptamer (i-DNA, i-RNA noma i-XNA)

Ama-Aptamers avame ukubhekisela kuma-aptamers e-nucleic acid. Ama-aptamers e-nucleic acid afaka ama-aptamers e-DNA, ama-aptamers e-RNA, kanye nama-aptamers e-XNA angama-aptamers e-nucleic acid aguqulwe ngamakhemikhali. Indlela ye-SELEX isetshenziswa kabanzi ekuthuthukiseni ama-aptamers. Ukuhamba komsebenzi okuyisisekelo kwezinsizakalo zokuthuthukisa ama-aptamer kufaka phakathi ukwakhiwa komtapo wolwazi, ukubopha okuqondiwe, ukuhlukaniswa kanye nokuhlanzwa, ukukhulisa, imijikelezo eminingi yokuhlola, kanye nokuhlonza ukulandelana. Sekuyiminyaka eminingi, sigxile ekwakheni umtapo wolwazi kanye nokuhlangenwe nakho okucebile ekuthuthukisweni kwama-aptamer. Sizibophezele ekuhlinzekeni amakhasimende ngezinsizakalo ezingcono.

Inqubo Yobuchwepheshe be-SELEX

Inqubo ye-SELEX inemijikelezo eminingi, ngayinye equkethe izinyathelo ezilandelayo ezibalulekile:

Ukubopha Umtapo Wolwazi kanye Nezinhloso

Umtapo wolwazi we-nucleic acid owakhiwe uxutshwa nama-molecule athile aqondiwe (njengamaprotheni, ama-molecule amancane ahlanganisiwe, njll.), ukuze ukulandelana kwe-nucleic acid kulabhulali kube nethuba lokubopha kuma-molecule aqondiwe.

Ukuhlukaniswa Kwama-Molecule Angaboshiwe

Ukulandelana kwe-nucleic acid okungaboshelwe ku-molecule okuqondiwe kuhlukaniswa nengxube ngezindlela ezithile ezifana ne-affinity chromatography, ukuhlukaniswa kwama-magnetic bead, njll.

Ukwandiswa Kwama-Molecule Ahlanganisayo

Uchungechunge lwe-nucleic acid oluboshwe ku-molecule eqondiwe luyandiswa, ngokuvamile kusetshenziswa ubuchwepheshe be-polymerase chain reaction (PCR). Esigabeni sokuhlola esilandelayo, uchungechunge olundisiwe luzosetshenziswa njengomtapo wolwazi wokuqala.

Umfanekiso 1: Inqubo yokuhlola ye-SELEX

Ipulatifomu Yokuhlola i-Aptamer

Isevisi yokuhlola i-Aptamer

I-Alpha Lifetech inikeza uhla oluhlukahlukene lwezinsizakalo zokuhlola i-aptamer ezikhethekile zisebenzisa izindlela ezahlukahlukene ze-SELEX zezinhlobo ezahlukene zama-molecule akho:

Izinhlobo Eziqondiwe	Imininingwane Yobuchwepheshe
Ukuhlolwa kwe-Aptamer yamaProtheyini yi-SELEX	Inhloso eyinhloko yokuhlolwa kwe-protein aptamer ukuhlola ama-aptamer angabopha ngqo kuma-molecule amaprotheni aqondiwe. Lawa ma-aptamer alula ukuwahlanganisa, azinzile futhi awathinteki kalula yizici zemvelo.
Ukuhlolwa kwe-Peptide Aptamer yi-SELEX	Ama-aptamer e-peptide ayisigaba semigqa emifushane ye-peptide enobungcweti obuphezulu kanye nobudlelwano, engabopha ngqo ezintweni eziqondiwe futhi ibonise ububanzi obukhulu bamandla okusetshenziswa emkhakheni wezokwelapha. Ngenqubo ethile yokuhlola, ama-aptamer e-peptide angabopha ngqo kuma-molecule aqondiwe ahlolwa kusuka emitatsheni eminingi yemigqa yokulandelana kwe-peptide engahleliwe.
Ukuhlolwa kwe-Aptamer yeseli ethile (i-Cell-SELEX)	Amaseli aqondiwe noma ama-molecule athile ebusweni beseli alungiswa njengezinhloso. Ama-Target angaba amaseli aphelele, ama-receptor kulwelwesi lweseli, amaprotheni, noma amanye ama-molecule amancane.
Ukuhlolwa kwe-Aptamer ye-Molecule Encane yi-Capture SELEX	I-Capture SELEX iyindlela yokuhlola ngaphakathi kwe-vitro yokuhlola ama-aptamer amancane e-molecule, okuwuhlobo lwe-SELEX. I-Capture SELEX ifaneleka kakhulu ekuhlolweni kwama-aptamer ama-target amancane e-molecule, ngokuvamile anamaqembu ambalwa asebenzayo futhi kunzima ukuwavimba ngqo ezisekelweni zesigaba esiqinile.
Izinsizakalo ze-SELEX ezisekelwe ezilwaneni ezibukhoma	Insizakalo yokuhlola izilwane eziphilayo iyindlela yokuhlola esetshenziswa kabanzi emikhakheni yesayensi yezinto eziphilayo, yezokwelapha kanye ne-biotechnology, esebenzisa izilwane eziphilayo njengemodeli yokuhlola ukuhlola nokuhlola ama-molecule athile, udokotela, ezokwelapha noma izinqubo zezinto eziphilayo. Lezi zinsizakalo zenzelwe ukulingisa indawo yemvelo emzimbeni womuntu ukuze zibikezele ngokunembile futhi zihlole ukusebenza kahle kanye nokuphepha kwemiphumela yokuhlola emzimbeni womuntu.

Isevisi yokwenza ngcono i-Aptamer

Ukuthambekela kwamanzi, ukulahlekelwa okukhulu kwe-affinity ngesikhathi sokukhiqizwa, kanye nokukhishwa okusheshayo kwama-aptamer kunciphisa ukusetshenziswa kwawo. Njengamanje, kuye kwahlolwa izindlela ezahlukahlukene zokwenza ngcono ukuze kuthuthukiswe ukusebenza kwama-aptamer.

Siphinde sibe nezindlela ezahlukahlukene zokwenza ngcono i-aptamer equkethe ukunqunywa, ukuguqulwa, ukuhlanganiswa neqembu elifanele (i-thiol, i-carboxy, i-amine, i-fluorophore, njll.).

Insizakalo yokuhlaziya izici ze-Aptamer

Insizakalo yokuhlaziya izici ze-Aptamer ibhekisela kwinsizakalo yobungcweti yokuxazulula isakhiwo sokuhlola ukusebenza kanye nokuqinisekiswa kokusebenza kwe-aptamer etholakele ukuqinisekisa ukuthi i-aptamer ihlangabezana nekhono elithile lokubopha, ukuzinza kanye nezidingo zokucacisa. Ihlanganisa kakhulu ukuhlaziywa kokuhambisana kanye nokucacisa, ukuhlolwa kokuzinza kanye nokuqinisekiswa komsebenzi webhayoloji.