Ka lawelawe nānā ʻana i ka waihona puke hōʻikeʻike Phage

Ua komo nui ʻo Alpha Lifetech i ka ʻenehana hōʻike Phage no nā makahiki he nui. Ua kūkulu ʻo ia i kahi kahua ʻenehana hōʻike Phage paʻa kūpono e mālama ana i ka manawa no ka noiʻi ʻepekema a i ʻole ka noiʻi papahana a hoʻomaʻalahi i ka hana ʻana ma hope. Hiki iā Alpha Lifetech ke hāʻawi i nā mea kūʻai aku me nā lawelawe e like me ka hana antibody vhh, ka hana antibody scFv, ka hana Fab Antibody.

Loaʻa iā Alpha Lifetech kahi kahua hōʻikeʻike phage M13/T7 piha e hōʻoia i ke kūkulu ʻana a me ka hana ʻana o nā hale waihona puke antibody. Hiki iā mākou ke hāʻawi i nā lawelawe i hana kūikawā ʻia, e like me ka hana ʻana o ka antibody scFv a me ka nānā ʻana o ka antibody kiʻekiʻe, e hoʻokō i kāu mau pono.

Ke Kaʻina Hana Kūkulu Hale Waihona Puke Hōʻikeʻike Phage

ʻO ke kaʻina hana o ke kūkulu ʻana i ka waihona puke phage penei: ua hoʻolālā ʻia nā primers kikoʻī no ka hoʻonui ʻana o ka PCR e loaʻa ai nā huahana, i hoʻopaʻa ʻia me ka enzyme me ka vector phage T7/M13, a ua kūkulu pono ʻia ka plasmid phage recombinant. Ua hoʻololi ʻia ka plasmid phage recombinant i nā cell receptor TG1, a laila ua uhi ʻia ma luna o kahi medium e loaʻa ana nā antibiotics kūpono, a ua hana ʻia ka moʻomeheu i hoʻonui ʻia ma hope o ka nānā ʻana i nā mea hoʻololi. Ma hope o nā pōʻai he nui o ka moʻomeheu, hana hou ka phage i nā manawa hana hou i loko o ka bacteria a hōʻike maikaʻi i ka protein target a i ʻole polypeptide. A laila ua hoʻomaʻemaʻe ʻia ka waihona puke phage e wehe i kekahi mau haumia a me nā phages i hoʻopaʻa ʻole ʻia.

Nā ʻAno Nānā o ka Hale Waihona Hōʻikeʻike Phage

ʻO ka holomua o ka hana ʻana o ka antibody hōʻike phage e hilinaʻi nui ʻia ana ma nā hale waihona hōʻike antibody phage, a ʻo nā ʻano maʻamau o ka phage panning he solid-phase a me ka liquid-phase screening. Hoʻonui ka Solid-phase antigen screening i nā antibodies kikoʻī ma o 3-4 mau puni o ka elution e loaʻa ai nā hale waihona hōʻike antibody phage; Hana ʻia ka wai-phase antigen screening ma ka hoʻoulu ʻana i nā waihona hōʻike biotin-labeled a me nā antibody phage a me ka hopu ʻana i nā phages i hoʻopaʻa ʻia me ka hoʻohana ʻana i nā beads magnetic. Hoʻopau ka Solid-phase screening i ka antigen hou aʻe a ua luku ʻia kekahi mau epitopes antigen. Hōʻike ʻia ka wai-phase screening e ka pono kiʻekiʻe a me ka hoʻonui ʻana, akā ʻaʻole kikoʻī loa nā antibodies i loaʻa. Aia ke ʻano o ka phage panning i nā pono hoʻokolohua, a e loaʻa kahi hopena i ka hopena o ka hana ʻana o ka phage display antibody.

Nānā ʻana i ka Antibody Throughput Kiʻekiʻe

Ua lilo ka hoʻomohala ʻana o ka antibody i mea wikiwiki i ka lāʻau lapaʻau hou. Aia nā ʻano hana hoʻomohala antibody like ʻole, akā hoʻohana nui ʻia ka nānā ʻana o ka antibody throughput kiʻekiʻe i ka ʻepekema lapaʻau. Hiki i ke kaʻina hana ke loaʻa ka nui o nā ʻano like ʻole o ka antibody repertoire. ʻO ka hui pū ʻana me nā ʻano hana nānā ʻana o ka waihona antibody kuʻuna hiki ke hoʻomaikaʻi i ka maikaʻi o ka nānā ʻana o nā waihona hōʻike phage. Ma hope o ka nānā ʻana o ka biology, hiki ke ʻike ʻia nā antibodies e ka ELISA haʻahaʻa a i ʻole ka nānā ʻana o ka antibody throughput kiʻekiʻe.

Kiʻi 1 Ka pakuhi kahe e wehewehe ana i ke kaʻina o nā hanana mai ke kūkulu ʻana i nā hale waihona puke antibody phage a hiki i ka hanauna o nā antibodies monoclonal me ka pilina kiʻekiʻe no nā antigens a me ka immunogenicity haʻahaʻa.(Kumu kuhikuhi: ʻO ka ʻenehana hōʻike Phage ma ke ʻano he kahua ikaika no ka ʻike ʻana i ka lāʻau lapaʻau Antibody - PMC (nih.gov))

Nā Mālama no ka Nānā ʻana i ka Hōʻikeʻike Phage AntibodyNā Mālama

Nā Manaʻo no ka Buffer

Pono ke koho ʻana o nā buffers e noʻonoʻo i ke kūpaʻa o ka pahuhopu, ka microenvironment nakinaki, a me ka liʻiliʻi o ka nakinaki ʻole kikoʻī i ke ʻano noi maoli. ʻO nā protein pili ʻole, nā mea holoi non-ionic, a me nā paʻakai physiological e kōkua i ka hōʻemi ʻana i ke kua. Aia kekahi mau kūlana ʻē aʻe, e like me kekahi mau pahuhopu e koi ai i ka hoʻohui ʻana o nā cations bivalent a i ʻole nā cofactors ʻē aʻe i ka buffer, hiki ke hoʻopaʻa pū ʻia nā cofactors protein me ka pahuhopu, hiki ke hoʻohui ʻia i ka buffer hoʻokaʻawale, a hiki ke hoʻohana ʻia ma ke ʻano he ala e hopu ai i ka pahuhopu.

Nā Manaʻo no ke Kaomi

ʻO nā noʻonoʻo kaomi koho maʻamau e pili ana i ka nui o nā molekala pahuhopu, ka manawa hoʻopaʻa, ka ikaika holoi, a pēlā aku. ʻO nā ʻano hana ʻē aʻe no ka hoʻonui ʻana i ke kaomi koho e pili ana i ka hoʻohana ʻana i nā denaturants (e like me nā waikawa, urea, a me guanidine), nā mea hoʻoheheʻe organik, ka hoʻonui ʻana i ka mahana, a i ʻole ka hoʻonui ʻana i ka nui o nā ligands hoʻokūkū a i ʻole nā mea i loko o ka buffer holoi.
I waena o lākou, he mea nui ka pōʻai mua o ka nānā ʻana. ʻO ka pōʻai mua o ka nānā ʻana, ʻo ia ka hopu ʻana i nā clones maikaʻi he nui e like me ka hiki mai ka waihona puke i kūkulu ʻia me ka wehe ʻana i nā clones kikoʻī ʻole. Pono e hoʻohana ʻia nā pahuhopu i uhi ʻia me ka porous a i ʻole ka nui o nā pahuhopu hiki ke hoʻoheheʻe ʻia e hōʻoia i ka hopu ʻia ʻana o ka nui o nā phages i hoʻopaʻa ʻia e mālama i ka ʻokoʻa o ka waihona puke. I nā pōʻai hope o ka nānā ʻana, pono e hoʻonui ʻia ke kaomi koho me ka hoʻonui ʻana o ka hoʻonui ʻana, ka holoi pinepine ʻana, a me ka hoʻonui ʻana i ka manawa holoi, hiki ke nānā ʻia nā clones me ka pilina kiʻekiʻe.

Hoʻolālā Nānā Maikaʻi ʻole

ʻO nā lepili antigen a me nā mea lawe ke kaʻina hana nānā ʻana no nā mea āpau ma nā hui antigen, e like me nā lepili, nā protein fusion, a me nā substrates kākoʻo. Hiki i ka nānā maikaʻi ʻole o nā mea i hoʻopaʻa ʻole ʻia ke kaupalena i ka hoʻonui ʻia ʻana o nā antibodies ʻē aʻe ma mua o ka antigen i hoʻopaʻa ʻia.
Hiki ke nānā pono ʻole ʻia nā pūnaewele holoʻokoʻa, nā liposomes, nā disc nano-phospholipid, a me nā VLP me ka hoʻohana ʻana i nā pūnaewele i hoʻololi ʻia a i ʻole nā pūnaewele i hoʻololi ʻole ʻia i ka wā e nānā ai i nā laina pūnaewele i hoʻololi ʻia.
Ka wehe ʻana i nā hui antigen paʻakikī Hiki ke hana ʻia ka nānā maikaʻi ʻole no nā molekala pahuhopu i ʻike ʻole ʻia a paʻakikī paha, e like me nā pūnaewele holoʻokoʻa a i ʻole nā protein haumia.
ʻO ke ʻano o nā antibodies e kūʻē i nā wahi kikoʻī o nā antigens, ʻo kekahi e hoʻokaʻawale i ka antibody i hiki ke hoʻokūkū me ka ligand ma ka hoʻohui ʻana i kahi kiʻekiʻe o nā ligands, a ʻo kekahi e ālai i ka antibody.

Ka Hoʻohana ʻana o ka Hōʻike Phage

*Ua lilo ka ʻike ʻana i ka antibody i mea nui i ka lāʻau lapaʻau hou. Aia nā ʻano ʻike antibody like ʻole, akā ua hoʻohana nui ʻia ka ʻenehana hōʻike phage i ka ʻepekema lapaʻau. Mai ka makahiki 1990, ua hoʻohana ʻia nā ʻano antibody like ʻole e kūkulu i nā hale waihona puke phage, me VHs, VHHs, scFvs, diabodies, a me Fab antibodies.
*ʻAe nā hale waihona puke peptide Phage i ka hoʻoholo wikiwiki ʻana i ke kaʻina o nā epitopes protein a ua lilo i mea hana ikaika no ka noiʻi ʻana i ka pilina ma waena o nā epitopes a me nā mea loaʻa antigen
*Hoʻohui ʻia nā ʻāpana antibody i ka G3P o ka M13 phage, a ma ke cloning ʻana i nā helu he nui o nā genes e hoʻopili ana i kahi ʻāpana antibody, hiki ke hoʻopuka ʻia nā waihona antibody hōʻike phage nui kahi e hiki ai ke koho ʻia nā antibodies like ʻole.
*He nui nā pono o ka ʻōnaehana hōʻike phage T7, me ka maʻalahi, ka palekana kiʻekiʻe, ke kūpaʻa, ka maʻalahi o ka mālama ʻana, a me ka lawe ʻana, no laila hoʻohana ʻia ka ʻōnaehana i nā lāʻau pale a me nā lāʻau lapaʻau.
*Hiki i ka ʻōnaehana hōʻike phage T7 ke ʻike i nā antigens like ʻole, e like me nā antigens ʻili o nā microorganism pathogenic a me nā antigens cancer.

ʻO Phage

NO KE AHA E KOHO AI IĀ MĀKOU

Hiki iā Alpha Lifetech ke hāʻawi i nā mea kūʻai aku me nā lawelawe e like me ka hana ʻana o ka antibody vhh, ka hana ʻana o ka antibody scFv, ka hana ʻana o Fab Antibody.