He aha ka Antibody Engineering?

Me ke kōkua o ka ʻenehana antibody, ua hiki ke hoʻololi i ka nui mole, pharmacokinetics, immunogenicity, hoʻopaʻa paʻa, kikoʻī a me ka hana hopena o nā antibodies. Ma hope o ka synthesizing antibodies, ʻo ka hoʻopaʻa ʻana o nā antibodies i mea waiwai nui i ka maʻi maʻi a me ka mālama ʻana. Ma o ka ʻenehana antibody, hiki iā lākou ke hoʻokō i nā pono o ka lāʻau lapaʻau a me ka hoʻomohala mua ʻana o ka diagnostic.

ʻO ke kumu o ka ʻenehana antibody ʻo ia ka hoʻolālā ʻana a hana i nā hana kikoʻī a paʻa i hiki ʻole i nā antibodies kūlohelohe ke hoʻokō, e kau ana i ke kumu no ka hana ʻana i nā antibodies therapeutic.

ʻO Alpha Lifetech, me kāna ʻike papahana nui i ka ʻenekinia antibody, hiki ke hāʻawi i nā lawelawe antibody monoclonal a polyclonal maʻamau no nā ʻano he nui, a me ka phage hōʻike antibody hale waihona puke a me nā lawelawe screening. Hiki i ka Alpha Lifetech ke hāʻawi aku i nā mea kūʻai aku me nā antibodies biosimilar maikaʻi a me nā huahana protein recombinant, a me nā lawelawe e pili ana, e hana i nā antibodies kūpono, kikoʻī, a paʻa. Ma ka hoʻohana ʻana i ka antibody piha, nā kahua protein a me nā ʻōnaehana hōʻike phage, hāʻawi mākou i nā lawelawe e uhi ana i ka upstream a me lalo o ka hana antibody, me nā lawelawe ʻenehana e like me ka antibody humanization, antibody purification, antibody sequencing, a me ka hōʻoia antibody.

ʻO ka pae paionia o ka ʻenehana antibody e pili ana i ʻelua ʻenehana:

--ʻenehana DNA Recombinant

--ʻenehana Hybridoma

ʻO ka hoʻomohala wikiwiki o ka ʻenehana antibody e pili ana i ʻekolu mau ʻenehana koʻikoʻi:

--Gene cloning technology and polymerase chain reaction

--Protein expression: Hoʻopuka ʻia nā protein recombinant e nā ʻōnaehana hōʻike e like me ka hū, nā maʻi kino koʻokoʻo, a me nā mea kanu.

--Kamepiula kōkua kūkulu hoʻolālā

ʻO ka hanauna mua o nā antibodies i hoʻohana ʻia no ka hana ʻana i nā antibodies chimeric, kahi i hoʻopili ʻia ai ka ʻāpana ʻokoʻa o ka ʻiole monoclonal antibodies i ka ʻāina mau o nā molekole IgG kanaka. Ua hoʻololi ʻia ka ʻāpana hoʻopaʻa antigen (CDR) o ka lua-generation mouse monoclonal antibody i loko o ka IgG kanaka. Ma waho aʻe o ka ʻāina CDR, ʻo nā antibodies ʻē aʻe a pau he mau antibodies kanaka, a ua hoʻoikaika ʻia e pale aku i ka hoʻoulu ʻana i nā pane anti-mouse antibody (HAMA) i ka wā e hoʻohana ai i nā antibodies clone ʻiole no ka mālama ʻana i ke kanaka.

 

No ke kūkulu ʻana i kahi waihona hōʻikeʻike phage, ʻo ka hana mua ka loaʻa ʻana o nā genes encoding antibodies, hiki ke hoʻokaʻawale ʻia mai nā keʻena B o nā holoholona i hoʻopaʻa ʻia (ke kūkulu hale waihona immune), unuhi pololei ʻia mai nā holoholona non immunized (kūkulu hale waihona kūlohelohe), a i ʻole i hui pū ʻia i loko o ka vitro me nā ʻāpana gene antibody (kūkulu hale waihona synthetic). A laila, hoʻonui ʻia nā genes e PCR, hoʻokomo ʻia i loko o nā plasmids, a hōʻike ʻia i loko o nā ʻōnaehana hoʻokipa kūpono (hōʻike hū (maʻamau ʻo Pichia pastoris), hōʻike prokaryotic (maʻamau ʻo E. coli), hōʻike kelepona mammalian, hōʻike cell mea kanu, a me ka hōʻike ʻana o nā pepeke i loaʻa i nā maʻi maʻi koʻokoʻo). ʻO ka mea maʻamau ka ʻōnaehana hōʻike E. coli, kahi e hoʻopili ai i kahi kaʻina antibody encoding kikoʻī ma luna o ka phage a hoʻopili i kekahi o nā protein shell phage (pIII a i ʻole pVIII). ʻO ka hui pū ʻana o, A hōʻike ʻia ma ka ʻili o nā bacteriophages. ʻO ke kumu o kēia ʻenehana ke kūkulu ʻana i kahi waihona hōʻikeʻike phage, ʻoi aku ka maikaʻi ma mua o nā hale waihona puke kūlohelohe i hiki ke loaʻa iā ia kahi paʻa kikoʻī. A laila, nānā ʻia nā antibodies me ka kikoʻī antigen ma o ke kaʻina koho biological, hoʻopaʻa ʻia nā antigens target, holoi pinepine ʻia nā phages i hoʻopaʻa ʻole ʻia, a holoi ʻia nā phages paʻa no ka hoʻonui hou ʻana. Ma hope o ʻekolu a ʻoi aʻe paha mau pōʻai o ka hana hou ʻana, hoʻokaʻawale ʻia nā antibodies kiʻekiʻe kiʻekiʻe.

Fig 3: Hana ʻia a me ka nānā ʻana i ka hale waihona puke Antibody

Hiki ke hoʻohana ʻia ka ʻenehana DNA Recombinant e hana i nā ʻāpana antibody. Hiki ke hoʻokahe ʻia nā antibodies Fab i ka mua e ka protease ʻōpū e hana (Fab ') 2 mau ʻāpana, a laila e ʻeli ʻia e ka papain e hana i nā ʻāpana Fab pākahi. ʻO ka ʻāpana Fv he VH a me VL, ʻaʻohe paʻa paʻa ma muli o ka nele o nā paʻa disulfide. No laila, hoʻopili pū ʻia ʻo VH a me VL ma o kahi peptide pōkole o 15-20 amino acids e hana i kahi ʻāpana ʻāpana kaulahao hoʻololi (scFv) antibody me ke kaumaha molekala ma kahi o 25KDa.

ʻO ke aʻo ʻana i ke ʻano o ka antibody ma Camelidae (Camel, LIama, a me Alpaca) ua hōʻike ʻia he mau kaulahao kaumaha wale nō nā antibodies a ʻaʻohe kaulahao māmā, no laila ua kapa ʻia lākou he mau kaulahao kaumaha (hcAb). Ua kapa ʻia ka ʻāpana hoʻololi o nā ʻelepona kaulahao koʻikoʻi i nā antibodies domain hoʻokahi a i ʻole nanobodies a i ʻole VHH, me ka nui o 12-15 kDa. E like me nā monomers, ʻaʻohe o lākou mau paʻa disulfide a paʻa loa, me kahi pilina kiʻekiʻe loa no nā antigens.

Fig 5: Heavy Chain Antibody a me VHH/ Nanobody

Hoʻohana ka ʻōlelo manuahi kelepona i ka hōʻike ʻana o ka DNA kūlohelohe a i ʻole synthetic no ka hoʻokō ʻana i ka synthesis protein in vitro, maʻamau me ka hoʻohana ʻana i ka ʻōnaehana hōʻike E. coli. Hoʻopuka koke ʻo ia i nā protein a pale i ke kaumaha metabolic a me ka cytotoxic ma nā cell i ka wā e hana ai i nā protein recombinant nui i loko o vivo. Hiki iā ia ke hana i nā protein paʻakikī e hoʻohui, e like me nā mea paʻakikī e hoʻololi ma hope o ka unuhi ʻana a i ʻole synthesize i nā protein membrane.