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ʻOihana Noiʻi ʻo Aptamer

Wahi a nā pono loiloi like ʻole o nā mea kūʻai aku, loaʻa iā Alpha Lifetech nā ʻano hoʻolālā loiloi aptamer like ʻole no nā mea kūʻai aku e koho ai.

KĀHEA IĀ MĀ˚OU
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ʻOihana Noiʻi ʻo Aptamer

Hoʻolauna iā Aptamer

ʻO nā Aptamers he mau molekala nucleic acid kaula hoʻokahi (e like me ka DNA a i ʻole RNA) i nānā ʻia in vitro mai kahi waihona puke o nā oligonucleotides synthetic me nā kaʻina i hoʻokaʻawale ʻia ma ke ʻano random e kahi kaʻina hana i kapa ʻia ʻo SELEX screening, kahi e pili ana i nā pōʻai he nui o ke koho iterative. ʻO nā ʻāpana nui o nā lawelawe hoʻomohala aptamer nona ʻo Alpha Lifetech e komo pū me ke kūkulu ʻana i ka waihona puke aptamer, ka nānā ʻana o aptamer SELEX, ka hoʻonohonoho ʻana o aptamer SELEX, ka nānā ʻana o ka kaʻina aptamer a me nā loiloi aptamer ʻē aʻe.

Hoʻohui ka hoʻonohonoho ʻana o SELEX i ka nānā ʻana o ka aptamer SELEX a me ka hoʻonohonoho ʻana o ke alapine kiʻekiʻe. Ma o ka hoʻonohonoho ʻana o ka aptamer SELEX, hiki ke hoʻoholo pono ʻia ke kaʻina kikoʻī o ka aptamer i hoʻopaʻa ʻia i ka pahuhopu, e hāʻawi ana i ka ʻikepili kumu no ka nānā ʻana a me ka hoʻohana ʻana i ka kaʻina aptamer ma hope.

Ua hoʻohana nui ʻia nā Aptamers i ka noiʻi ʻepekema, ka mālama ʻana i nā maʻi, ke kūkulu ʻana o biosensor, ka ʻike ʻana i nā lāʻau lapaʻau a me ka nānā ʻana i ke kaiapuni. Eia nō naʻe, ua maʻalahi ka hoʻohaumia ʻana o nā aptamers oligonucleotide i nānā ʻia ma vitro i loko o vivo a hōʻike pū i ka ʻawahia. No laila, ma hope o ka hoʻomaikaʻi ʻana i nā aptamers i nānā ʻia, pono ka loiloi in vitro e loiloi i ka holomua o ka hoʻomohala ʻana o ka aptamer. Wahi a nā pono loiloi like ʻole o nā mea kūʻai aku, loaʻa iā Alpha Lifetech nā ʻano hana loiloi aptamer like ʻole no nā mea kūʻai aku e koho ai.
loiloi aptamer
Kiʻi 1 Kiʻikuhi o ka loiloi aptamer. Kumu kuhikuhi:ʻO Thevendran R, Citartan M. 2022. Nā hoʻāʻo e kuhi i ka pilina paʻa o nā Aptamers.

Hoʻolauna i ka lawelawe noiʻi Aptamer

Ka Nānā Paʻa ʻana o Aptamer

Hoʻokolohua hoʻohaʻahaʻa nuclease
Ma ka hoʻoulu ʻana i ka aptamer me nā ʻano nucleases like ʻole (e like me DNase, RNase, a pēlā aku) ma lalo o nā kūlana kikoʻī, ʻike ʻia ka hōʻino ʻia o ka aptamer, i mea e loiloi ai i kona hiki ke pale i ka hōʻino ʻia. He mea maikaʻi kēia no ka hoʻoholo ʻana i ke kūpaʻa a me ka lōʻihi o ka aptamer i nā hana maoli.
Ke ʻano hana lepili fluorescent
Ua loiloi ʻia ke kūpaʻa o ka aptamer ma ke ʻano ʻē ma ka lepili ʻana i nā fluorophores ma ka aptamer a me ka nānā ʻana i nā loli o ka hōʻailona fluorescence ma mua a ma hope o ka nakinaki ʻana me ka pahuhopu. No ka laʻana, ke nakinaki kahi aptamer i kahi pahuhopu, hiki ke loli kona ʻano, e hopena ana i kahi hōʻailona fluorescence i hoʻonui ʻia a emi paha.
Ka loiloi kūpaʻa wela
Hoʻoholo ʻia ke kūpaʻa wela o ka aptamer ma ke ana ʻana i ka loli o kona mahana hoʻoheheʻe (waiwai Tm) ma nā mahana like ʻole. ʻO ke kiʻekiʻe o ka mahana hoʻoheheʻe o ka aptamer, ʻoi aku ka maikaʻi o kona kūpaʻa wela.
Ka loiloi kūpaʻa hoʻololi
Ua hoʻohana ʻia ka Surface plasmon resonance (SPR) a me nā ʻenehana ʻē aʻe e nānā i ke kaʻina hana hoʻopaʻa a me ka decoupling ma waena o ka aptamer a me ka pahuhopu i ka manawa maoli, loaʻa nā palena dynamic (e like me ka binding rate constant, dissociation rate constant, etc.), e hoʻomaopopo hou i ke ʻano hana pilina ma waena o ka aptamer a me ka pahuhopu, a loiloi i kona kūpaʻa dynamic.
Ka loiloi kūpaʻa kūkulu
Ua hoʻohana ʻia ka crystallography X-ray, ka resonance magnetic nuclear (NMR) a me nā ʻano hana biology ʻē aʻe e kālailai i ke ʻano ʻekolu-dimensional o ka aptamer a me kona kūpaʻa kūkulu.

Ka Nānā Kūikawā ʻo Aptamer

Hoʻāʻo hoʻopaʻa ʻana o Aptamer
Hoʻokolokolo ʻia ka affinity ma ka hoʻokō ʻana i ka assay assay a aptamer e ana i ke kūpaʻa paʻa ma waena o ka aptamer a me ka mole i manaʻo ʻia, e like me ke kūpaʻa dissociation Kd. ʻO ka waiwai Kd haʻahaʻa e hōʻike ana i kahi affinity kiʻekiʻe. Hiki i ka assay a Aptamer binding ke nānā i nā moho lāʻau lapaʻau me ka hiki ke hoʻopaʻa kiʻekiʻe, a laila e hana i nā haʻawina pharmacodynamics a me pharmacokinetics ma hope.
Hoʻokolohua loiloi hoʻohuli
ʻO ka hoʻāʻo pale hope he ʻano pale ia e kāpae koke i nā molekala ʻaʻole i hoʻolālā ʻia mai kahi nui o nā molekala moho. Hoʻolālā ʻia ka pale hope e kāpae i nā molekala ʻaʻohe ona waiwai a hana paha. ʻO ke kī i ka pale hope, ʻo ia ke koho ʻana i nā māka pale hope kūpono a i ʻole nā ​​kūlana e hiki ai ke ʻike a wehe ʻia nā molekala ʻaʻole i hoʻolālā ʻia i ka wā o ke kaʻina hana pale. Ma kēia hoʻokolohua, he mea nui loa ke koho ʻana i nā molekala ʻaʻole i hoʻolālā ʻia. Pono e hōʻoia ʻia he mau mea hōʻike nā molekala ʻaʻole i hoʻolālā ʻia i koho ʻia no nā mea e hoʻopilikia paha i ka pale ʻana o ka aptamer a ua uhi piha ʻia nā mea hoʻopilikia e like me ka hiki.
Hoʻokolohua kāohi hoʻokūkū
Ma ka hoʻohui ʻana i nā mea hoʻokūkū nui loa (e like me nā molekala me ke ʻano like me ka molekala i manaʻo ʻia) i ka ʻōnaehana hoʻopaʻa o ka aptamer a me ka molekala i manaʻo ʻia, ʻike ʻia ka loli o ka hiki ke hoʻopaʻa o ka aptamer a me ka molekala i manaʻo ʻia. Inā ua emi nui ka hiki o ka aptamer ke hoʻopaʻa i ka molekala i manaʻo ʻia, hōʻike ia he kiʻekiʻe ke kikoʻī o ka aptamer.
I kekahi mau hihia, hiki ke hoʻohana ʻia nā hoʻokolohua kāohi hoʻokūkū ma ke ʻano he ʻāpana a i ʻole he mea hoʻohui i nā hoʻokolohua nānā hope. No ka laʻana, i ke kaʻina hana nānā lāʻau lapaʻau, hiki ke loiloi ʻia ka hiki o ka molekole lāʻau lapaʻau ke hoʻopaʻa i ka protein i manaʻo ʻia e nā hoʻokolohua kāohi hoʻokūkū, a laila hiki ke hoʻohana ʻia nā hoʻokolohua nānā hope e wehe i kēlā mau hui i hoʻopaʻa ikaika ʻia i nā cell maʻamau a i ʻole nā ​​​​ʻiʻo.

Ka Nānā ʻana o ka Cytotoxicity Aptamer

Aia kekahi mau ʻano hoʻokolohua like ʻole no ka nānā ʻana i ka aptamer cytotoxicity, i hoʻolālā ʻia e loiloi i nā hopena o nā aptamers ma ke ola ʻana o nā cell, ka hoʻonui ʻana, a i ʻole ka hana.
Nā ʻAno Hana Hoʻolauna kikoʻī Pōmaikaʻi Ka hemahema
Ke ʻAno ʻIke MTT ʻO ka hoʻāʻo MTT kahi ʻano hana i hoʻokumu ʻia ma ka hana enzyme i loko o ka mitochondria o nā cell ola. Hiki i ka succinate dehydrogenase i loko o ka mitochondria o nā cell ola ke hōʻemi i ka MTT exogenous i ka kristal polū-poni wai ʻole Formazan a waiho i loko o ke kelepona, ʻoiai ʻaʻohe hana o nā cell make. ʻO ka hoʻoheheʻe ʻana i kēia mau kristal e ka dimethyl sulfoxide (DMSO) a me ka ʻike ʻana i ka absorbance ma nā nalu kikoʻī (e like me 490nm a i ʻole 570nm) ma kahi enzymoleter hiki ke hōʻike pololei ʻole i ka helu o nā cell ola a no laila e loiloi i ka cytotoxicity o ka aptamer. ʻO ke kūlana kiʻekiʻe, ka hoʻokele waiwai a me ka maʻalahi ʻO ka ukana hana kaumaha, hiki i nā mea hoʻoheheʻe organik ke hōʻino i nā cell; Hiki ke loaʻa nā hopena hoʻokolohua hope loa, a ʻaʻole hiki ke nānā ʻia ke kaʻina holoʻokoʻa o ka cytotoxicity
Ke ʻAno ʻIke ʻana iā CCK-8 ʻO ka Cell Counting Kit-8 (CCK-8) kahi ʻano hana ʻike colorimetric kiʻekiʻe-ʻike kiʻekiʻe, ʻaʻole radioactive. Loaʻa iā CCK-8 ka WST-8, kahi i hoʻemi ʻia e ka dehydrogenase i loko o ka mitochondria o nā cell ola e hana i ka wahie methyl zan ʻalani hiki ke hoʻoheheʻe ʻia i ka wai. ʻO ka nui o ka mezan dye i hana ʻia e pili pono ana i ka helu o nā cell ola, a hiki ke hōʻike pololei ʻole ʻia ka helu o nā cell ola ma ke ana ʻana i ka waiwai omo mālamalama o ka Mezan dye ma ka nalu o 450nm, i mea e loiloi ai i ka cytotoxicity o ka aptamer. Hana maʻalahi, ʻaʻohe pono e holoi i nā pūnaewele, ʻike wikiwiki, ākea ka laulā ʻike linear, ʻike kiʻekiʻe, hana hou maikaʻi, liʻiliʻi ka cytotoxicity Kumukūʻai reagent kiʻekiʻe; He mea paʻakikī i kekahi manawa ke haʻi aku inā ʻo ka emi ʻana o ka absorbance ma muli o ka emi ʻana o ka helu o nā cell ola a i ʻole ka emi ʻana o ka hana o ka cell dehydrogenase ponoʻī
Ke ʻAno ʻIke LDH ʻO LDH (lactate dehydrogenase) kahi enzyme i paʻa i loko o ka cytoplasm o nā cell, a i ka wā e hōʻino ʻia ai ka membrane cell, hoʻokuʻu ʻia ʻo LDH ma waho o ke cell. Hiki i ka LDH ke hoʻoulu i ka waikawa lactic e hana i ka pyruvate, a hana pū me INT (tetrazolium salts) e hana i ka mea crystalline poni. Ma ke ana ʻana i kona absorbance ma kahi nalu kikoʻī (e like me 490nm), hiki ke hōʻike ʻia ke kekelē o ka hōʻino ʻana o ke cell, a laila hiki ke loiloi ʻia ka cytotoxicity o ka aptamer. Hōʻike pololei ia i ka make ʻana o nā cell, a emi iho ka hōʻino ʻana i nā cell, a ʻaʻohe haumia isotope radioactive ʻO ka pono e lawe pinepine i nā cell mai loko mai o ka incubator, ʻoi aku ka paʻakikī o ka hana; Hiki ke loaʻa nā hopena hoʻokolohua hope loa, a ʻaʻole hiki ke nānā ʻia ke kaʻina hana piha o ka cytotoxicity
Ka Nānā ʻana i ke Kiʻi ʻana o nā Kelepona Ola i ka Manawa Maoli Ma ka hoʻohana ʻana i ka mea kālailai kiʻi kelepona ola manawa maoli, ua kau ʻia ka mea hana i loko o ka incubator e nānā a hoʻopaʻa i ke kaʻina holoʻokoʻa o ke kaʻina ulu ʻana o ke kelepona i ka manawa maoli. Hiki ke loiloi ʻia ka cytotoxicity o nā aptamers ma ka nānā ʻana i nā loli morphological a me nā piʻo ulu o nā kelepona. Hiki i kēia ʻano hana ke hāʻawi i ka wikiō a me nā hopena i helu ʻia o nā kaʻina hana cytotoxic me ka mālama ʻana i ke kaiapuni ulu kelepona paʻa. Hiki iā ia ke nānā i ke kaʻina hana cytotoxic i ka manawa maoli, nā kiʻi ʻole luku, e hōʻemi i ka hoʻopilikia a me ka hōʻino ʻana i nā cell; Loaʻa nā hopena wikiō a me nā hopena i helu ʻia no ka loiloi hohonu. He kiʻekiʻe ke kumukūʻai o nā lako a koi aku i nā mākau hana loea a me ka hiki ke kālailai ʻikepili

KACHI

Ka Pōmaikaʻi o ka Lawelawe Noiʻi Aptamer

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Ka Mana Hoʻomalu Kūlana Holoʻokoʻa

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Nā Aptamers i hoʻomaikaʻi ʻia me ke kūpaʻa kiʻekiʻe

ikona08

Hoʻokele waiwai a me ka pono. ʻOi aku ka hoʻokūkū o ke kumukūʻai, ʻoi aku ka maikaʻi o ka lawelawe

ikona10

Nā wahi noiʻi piha a me nā ʻano noiʻi he nui

ikona09

Nā lawelawe kūkākūkā mua-kūʻai aku a me ke kākoʻo ma hope o ke kūʻai aku

ikona07

Ka ʻike waiwai i ka nānā ʻana o ka aptamer

ikona05-1

Hui R&D kiʻekiʻe

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Nā lawelawe pili aptamer i hoʻopilikino ʻia

E hoʻi i ka ʻAoʻao Paepae Hoʻomohala Aptamer

Inā he mau nīnau kāu, e ʻoluʻolu e hoʻokaʻaʻike mai iā mākou i kēlā me kēia manawa.

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