Whakataki Ratonga Whakaaturanga Phage

Ko te whakaaturanga phage, hei tikanga ngota ngota e ahu mai ana i te whakarerekētanga ira o te DNA phage, kua noho hei taputapu kaha ki te whakaputa i ngā whakamātautau ngota ngota ki ngā ūnga motuhake mā te whakaatu i ēnei kongakonga ki te mata o te phage, ā, nō reira ka tīpakohia ngā kongakonga peptide/antibody me ngā āhuatanga here motuhake mai i te tini o ngā momo (whare pukapuka).

Tekau tau te roa o te wheako o Alpha Lifetech ki te whakawhanake i ngā paturopi whakaatu phage, ā, ka taea e rātou te whakarato ratonga hanga whare pukapuka paturopi ngaio me te tirotiro mō ā mātou kiritaki. E ai ki ngā hiahia o ngā kiritaki, ka taea e mātou te whakarato i ngā momo whare pukapuka paturopi rerekē, pērā i ngā whare pukapuka ārai mate, ngā whare pukapuka taketake, ngā whare pukapuka hangai me ngā whare pukapuka haurua-hangai. Ko te hangarau whare pukapuka whakaatu phage, e whakakotahi ana i ngā ira paturopi me ngā ira o ngā pūmua uhi phage pērā i te M13, koia te hangarau whakarite paturopi recombinant tino noa, ā, e whakamahia whānuitia ana i roto i te whakawhanaketanga o ngā paturopi tino motuhake.

Ka werohia e Alpha Lifetech ngā arapaka, ngā rāma, ngā arapaka, me ngā mango, me ētahi atu, kia eke ai tō rātou whai huatanga ki te 10^5. Ka tukuna atu e mātou te pūrongo whakamātautau whai huatanga ki te kiritaki hei whakarite i te tika me te pono o ngā raraunga. He iti te paitini me te iti o te ārai mate kei roto i ngā whare pukapuka taketake. He maha ngā whare pukapuka paturopi taketake kua oti te hanga a Alpha Lifetech, ka taea te whakamahi tika mō te tirotiro paturopi me te kore e werohia ngā kararehe, ā, ka whakapotohia te wā o te kaupapa, ka whakatere i te ahunga whakamua o te whakamātautau a te kiritaki, ā, ka whakamanahia te pono o te mahi a te paturopi.

Pikitia 1: Te kaupapa whakaatu phage

Tā Mātou Pūnaha Whakaaturanga Phage

Kei te Ripanga 1 ngā mōhiohio matua mō te pūnaha whakaatu phage M13/T4/T7.

Ripanga 1 Ngā waka whakaatu phage rerekē me ō rātou āhuatanga
	M13	T4	T7
Rahi o te Ira	6407 bp	168895 bp	39937 bp
Whakaatu Pūmua	pVI, pIII me te pVIII	SOC me HOC	gp10B
Rahi Whakaatu	>110 kDa ko te pIII
Rahi Whakaatu
Te Kīanga Whakaatu
Te Kīanga Whakaatu
Huringa Ora	Risogene	Lytic	Lytic

Ka taea e Alpha Lfetech te whakarato

Ka whakakotahi a Phage Display i te iraira me te āhuahanga ki te kotahi, ka whakakotahi i te whiriwhiri me te whakanui me ngā āheinga tirotiro kaha. Ka taea e Alpha Lifetech te whakarato i te whānuitanga o ngā ratonga mō te whakawhanaketanga o te paturopi whakaaturanga phage.
Ko ngā ratonga matua ko: te tūāpapa whare pukapuka paturopi VHH, te tūāpapa whare pukapuka paturopi scFv, te tūāpapa whare pukapuka paturopi Fab, te tūāpapa hanga whare pukapuka phage, te tūāpapa tirotiro whare pukapuka phage me te ratonga whakatangata paturopi me ētahi atu ratonga.

te whakawhanaketanga o te paturopi-Alpha Lifetech

Pikitia 2: Te tukanga ratonga whakawhanake whakaaturanga Phage

Te Tukanga Whakawhanaketanga Paturopi Whakaaturanga Phage

Ka taea e mātou te wero i ngā kararehe ki ngā pūmua i whakaatuhia i roto i tā mātou taiwhanga, i tukuna mai rānei e ā mātou kiritaki, tae atu ki ngā momo pūmua macromolecular, peptides me ngā pūmua membrane. Mā te whakamātautau whai huatanga ELISA, i whakahaerehia e ā mātou kairangahau te wehenga PBMC me te tango RNA mai i te toto o ngā alpaca me ngā hua wero pai rawa atu. Mā te whakawhiti whakamuri me te whakanui ira, i hangaia e mātou te ira ki runga i ngā kawe phage pMESC, pComb3XSS, pCANTAB 5E rānei. Mā te whakawhiti hiko o ngā pūtau TG1 E. coli, ka taea te whiwhi i ngā whare pukapuka paturopi phage me te kaha whare pukapuka nui atu i te 10^9. He pai te kanorau o ngā whare pukapuka paturopi me te kaha whare pukapuka nui, ā, ka āwhina i te kimi paturopi he kaha te hononga. Hei whakarite i te kounga o te whare pukapuka paturopi, ka whakaahuahia e mātou te kaha whare pukapuka me te tere whakauru o te whare pukapuka. Ka raupapahia e mātou ngā whare pukapuka hei whakarato i ā mātou kiritaki ki ngā raupapatanga 30-50 e hiahiatia ana. Kātahi ka whakahaerehia e mātou te tirotiro whare pukapuka whakaaturanga phage. Tuatahi, ka herea te antigen ki runga i ngā pereti iti polypropylene, ā, mā roto i ngā huringa tirotiro e 3-5, ka tangohia ngā paturopi phage he ngoikore te kaha ki te here, ā, ka puritia ngā ira motuhake e herea ana ki te antigen. Mō te whakaaturanga phage, ka whakamahia te tikanga ELISA kia puta he ira pai, engari mō te whakaaturanga rewena, ka whakamahia te tikanga FACS kia whakaatuhia e ngā pūtau ngā kongakonga paturopi recombinant mā te whakamahi i ngā ira kua tohua. Mā te tirotiro, ka whakaritea e mātou ngā paturopi he tino aro, he tino motuhake, he tino kaha hoki.

Pikitia 3 Te tukanga hanga paturopi whakaaturanga phage

NGĀ PAINGA O TE Ratonga
Ngā painga o te RATONGA

He nui te whakapau kaha o Alpha Lifetech ki te hangarau whakaatu phage, ā, ka taea e ia te whakarato ratonga phage whānui ake, pai ake hoki ki ngā kiritaki.

Te wā me te whai huatanga teitei

Ka taea e tā mātou kamupene te rapu i ngā paturopi e pai ana i roto i te wā poto; ka taea e mātou te whakahaere i ngā whakamātautau tirotiro paturopi maha e ai ki ngā hiahia o ngā kiritaki.

Te kounga teitei o te hua

Ka āwhina tā mātou kamupene i ngā kiritaki ki te whakarite i ngā paturopi he tino motuhake me te pumau, ka whakapiki ake i te putanga o ngā hua rangahau pūtaiao.

Whakaratohia he whānuitanga o ngā kōwhiringa

Ka taea e tā mātou kamupene te hanga whare pukapuka paturopi maha-momo (tangata, kiore, rāpeti, ārapa, me ētahi atu) ā, ka taea hoki te hanga i ngā momo whare pukapuka paturopi maha (scfv, Fab, VHH, me ētahi atu).

Te kaha nui

He nui te kaha o tā mātou whare pukapuka paturopi, neke atu i te 10^9 paturopi.

Ngā Pātai Auau - Ngā raruraru whānui o te whakaaturanga phage

Ngā Pātai Whakaaturanga Pūnaha ◢

P.
He aha te hangarau whakaatu phage?

A.
Ko te hangarau whakaatu phage he whakauru i te ira ūnga ke ki te tūranga motuhake o te pūmua capsid phage, ka whakaatu i te ira ke ki te mata o te phage me te kore e pā ki te mahi noa o te phage. Whai muri i ētahi huringa tirotiro, i whiwhihia te pūmua ūnga he kaha te motuhake me te kaha koiora teitei.
P.
He aha ngā painga o te hangarau whakaaturanga phage?

A.
Ka taea te whakaputa i ngā whare pukapuka paturopi kaha-teitei mā te whakamahi i tēnei hangarau. He poto te huringa tirotiro, he māmā te tukanga whakahaere, ā, he whānui te whānuitanga o ngā tono, e āhei ai te tautuhi i ngā raupapatanga tino pai mō ngā peptide, ngā pūmua rānei kua tohua. I roto i ngā tono e pā ana ki te huaketo, ka taea e ngā kairangahau te kite i ngā paturopi e herea ana ki ngā epitope huaketo nui mā te tirotiro i ngā whare pukapuka paturopi nui. Ka taea e tēnei tukanga te aukati i te whakaurunga, te tāruarua, te karo rānei i ngā tikanga ārai mate o ngā huaketo, ā, mā reira ka whakatūria he turanga mō te maimoatanga mate.
P.
He aha te rerekētanga i waenga i te hangarau hybridoma me te hangarau phage?

A.
He whānui ake te whakamahinga o te hangarau whakaaturanga phage i te hangarau hybridoma, e iti ana ki te whakaputa i tētahi rahinga iti o ngā paturopi hei ārai i ngā momo ārai mate motuhake. He rerekē, ka taea e te hangarau whakaaturanga phage te whakaatu i tētahi whare pukapuka paturopi katoa i ahu mai i ngā momo momo ārai mate rerekē.
P.
He aha ngā whakamahinga o te hangarau whakaaturanga phage?

A.
He whānuitia ngā whakamahinga o te hangarau whakaatu phage, tae atu ki te tirotiro i ngā kaiwhakawhiwhi o ngā moroiti mate, te ako i ngā taunekeneke pūmua, te whai i te tuku tohu i waenga i ngā pūtau, te tautuhi i ngā mate kararehe, me te whakawhanake i ngā kano kano, me ētahi atu mara.
P.
He aha te tumanako mō te hangarau whakaatu phage?

A.
He maha ngā painga o te hangarau whakaaturanga phage, ā, ka taea te whakamahi puta noa i ngā momo mara. Heoi, ko te wero matua i roto i te whakawhanake i ngā paturopi recombinant mā te whakamahi i te tikanga whakaaturanga phage ko te uaua me te utu e pā ana ki te tukanga. Me whakauru ngā tikanga koiora ngota, tae atu ki te whakarerekētanga ira, te cloning, me te whakaaturanga, me ngā tātaritanga ārai mate, matū koiora, me te koiora ā-tinana hei tautuhi me te whakaahua i ngā paturopi e hiahiatia ana.

Ngā Pātai e pā ana ki te Antigen ◢

P.
He aha ngā tikanga e whakaatu ana i te antigen?

A.
He hononga tino nui te whakaaturanga o te antigen. Ko ngā tikanga o te whakaaturanga antigen ko te whakaaturanga tika o te antigen kua pania, te whakaaturanga ā-kore o te antigen kua pania, te whakaaturanga antigen mā te tirotiro pūtau katoa, te whakaaturanga antigen mā roto i ngā liposome, ngā kōpae nano-phospholipid me ngā VLP.
P.
He aha ngā rerekētanga i waenga i ngā momo whakaaturanga o te antigen?

A.
He tino māmā te tikanga paninga tika, engari he ngāwari te whakarerekē i te āhua o te pūmua ārai mate, ā, he uaua te mahi paninga ā-kore, engari ka taea te pupuri i te āhua o te pūmua ārai mate me te whakapai ake i te whai huatanga o te whakaaturanga. Ka taea e te tirotiro pūtau te whakatauira i te tirohanga tūturu i roto i te tinana ora, engari he raruraru kei te herea kore-motuhake. He iti te rahi o te kōpae nano-phospholipid, he pai te tuku me te pumau pai; ka honoa ngā VLP ki te pūmua ārai mate, ka takaihia rānei ki te pūmua ārai mate, he nui te kaha ārai mate me te haumaru pai, ā, ka taea te whakaoho i te urupare ārai mate kaha.
P.
He aha te pānga o te kukū o te antigen ki ngā hua tirotiro o te hangarau whakaaturanga phage?

A.
Ki te iti rawa te kukū, ka heke te tūponotanga o te herenga ki te phage, ka iti ake te whai huatanga o te tirotiro. I tetahi atu taha, mā te kukū tino tiketike ka piki ake te tūponotanga o te herenga kore-motuhake, ka whakanui ake i te tohu papamuri, ā, ka ārai i te tautuhi i ngā ira-rua pai.
P.
Me pēhea tātou e whakarite ai kia mau tonu te āhua tūturu o te antigen i te wā o te tukanga tātaritanga?

A.
Ka taea te whakamahi i te tikanga whakapiri ngawari hei karo i te pāmahana teitei, te waikawa kaha me te kawakore. I whakamahia te tikanga paninga ā-kore me te pūnaha biotin-streptavidin hei whakaiti i te kino ki te hanganga o te antigen. Mena he pūmua membrane te antigen, ka taea te whakaatu ki te mata o ngā pūtau kore e pakaru, ka taea rānei te whakamahi hei whakatauira i te taiao membrane, pērā i te nano-disk, hei pupuri i tōna āhua pūmua membrane tūturu.
P.
Me pēhea te manatoko i te motuhaketanga o te here o ngā paturopi i tirotirohia e te hangarau whakaatu phage ki ngā antigens?

A.
Haunga ngā whakamātautau noa pēnei i te ELISA, te Western Blotting (WB), me te Immunofluorescence (IF), ka taea te whakamahi i ngā tikanga kimi Surface Plasmon Resonance (SPR) me te Bio-Layer Interferometry (BLI) hei aroturuki i ngā tukanga here me te wetewete o ngā paturopi me ngā antigens i te wā tūturu, kia taea ai te whakatau i ngā tawhā hononga me te nekeneke.

Ngā Pātai e Pā Ana ki te Ārai Mate ◢

P.
He aha ngā urupare ārai mate whānui a ngā kararehe i roto i te hangarau whakaatu phage?

A.
Ka taea te whakamahi i te whakaaturanga phage hei whakaputa paturopi monoclonal tino hononga mō te whānuitanga o ngā momo whakaputa paturopi, tae atu ki te tangata, kiore, kiore iti, rāpeti, heihei, kamera, arapaka, kau, kuri, hipi, makimaki, me ngā mango.
P.
He aha ngā āhuatanga me ngā horopaki e tika ana mō ngā whare pukapuka paturopi i ahu mai i ngā momo kararehe rerekē i roto i te hangarau whakaatu phage?

A.
Kua tino mōhiotia te hangarau hanga o ngā whare pukapuka paturopi kiore, he iti te utu, ā, he poto te roa o te whakamātautau, he mea pai mō te tirotiro paturopi tuatahi me te rangahau taketake. He kaha te hononga me te motuhake o ngā paturopi rāpeti, me te whānuitanga o te mōhiotanga epitope ki te whakataurite ki ngā paturopi kiore. Ka taea te tiki tika i ngā paturopi tangata mai i ngā whare pukapuka paturopi tangata, kia kore ai e puta he urupare ārai mate ka puta mai i te wā e tukuna ana ngā paturopi heterologous ki te tinana tangata. He nui te uara o tēnei huarahi i roto i te rangahau me te whanaketanga rongoā, inā koa i roto i te hanganga o ngā paturopi rongoā.
P.
He aha te immunogen kararehe whānui e whakamahia ana i roto i te hangarau whakaaturanga phage?

A.
Ko ngā ārai mate kararehe ka wehea whānuitia ki te pūmua pūmua, te pūmua polypeptide, te pūmua waikawa karihi me te pūmua e pā ana ki te mate, ā, ko ētahi o ēnei ko te pūmua e pā ana ki te mate ko te pūmua huaketo (te matūriki huaketo katoa, te pūmua huaketo, te pūmua kore-hanganga huaketo, me ētahi atu), te pūmua kitakita (te tangohanga kitakita katoa, te wāhanga pakitara pūtau, te pūmua huna, te paitini, me ētahi atu) me te pūmua pirinoa.
P.
Me pēhea tā tātou e aro atu ai ki te take o te manawanui o te ārai mate i roto i ngā kararehe i te wā e whakautu ana te ārai mate?

A.
Whakatikatikahia ngā matū ārai mate mā te whakakapi i ngā matū ārai mate ki ngā pūmua hangarua me te whakakapi i ngā matūriki huaketo tinei ahi ki ngā pūmua hanganga o ngā huaketo. Whakatikatikahia te hoahoa whakamātautau me te whakarerekē i te horopeta ārai mate.
P.
Me pēhea te whakapai ake i te kaha ārai mate o ngā immunogens i roto i te hangarau whakaaturanga phage?

A.
Ka taea te whakanui ake i te kaha ārai mate mā te hono i ngā antigens ki ngā pūmua kawe, pērā i te albumin toto kau. Hei tāpiri, ka taea te whakamahi i ngā āwhina ārai mate, tae atu ki te āwhina Freund me te āwhina konumohe, hei whakaroa i te wā noho i roto i te tinana me te whakaoho i te whakahohenga me te tipu o ngā pūtau ārai mate, ā, mā reira ka whakapai ake i te kaha ārai mate.

Ngā Pātai Hanga Whare Pukapuka ◢

P.
He aha te whare pukapuka paturopi phage?

A.
Ko te whare pukapuka paturopi phage he kohinga o ngā paturopi phage kanorau i hangaia mā te whakakotahi i tētahi huinga katoa o ngā ira rohe rerekē mai i ngā paturopi ki ngā kawe phage me te whakaatu i aua ira ki runga i ngā mata o ngā phage. Ko te hua o tēnei tukanga he hanganga o tētahi whare pukapuka paturopi phage.
P.
Te tukanga hanga i tētahi whare pukapuka paturopi phage

A.
Ko te tukanga whānui o te whakaaturanga phage he maha ngā mahi matua: te tango i ngā pūtau mononuclear toto ā-taha (PBMC) mai i ngā kararehe kua werohia, te wehe i te RNA katoa me te whakawhiti ki te DNA tautoko (cDNA), te whakanui ake i te ira ūnga mā te whakamahi i te hangarau tauhohenga mekameka polymerase (PCR), te tārua i te ira ūnga ki roto i te vector phage, me te huri i te vector ki roto i te pūtau manaaki mō te whakaaturanga. Ko te hua o tēnei tukanga ko te hanganga o tētahi whare pukapuka whakaaturanga phage, ka taea te tautuhi tere i ngā paturopi tino hononga. Nō muri mai, ka taea e tēnei whare pukapuka te whakamahi hei rauemi nui mō te whakawhanake i ngā kano kano, ngā rongoā whakaora, me ngā matū tātaritanga.
P.
He aha ngā āhuatanga matua o te hanga i tētahi whare pukapuka kounga teitei?

A.

Me whakarite kia rawaka te tapeke o ngā PBMC. Me whakarite kia kounga te RNA me te kore e heke.

Hei whakarite i te tapeke o ngā whakawhiti, me 10 ki te 100 ngā wā o te kanorau ariā te kaha o te whare pukapuka.
P.
He aha ngā whakarōpūtanga o te whare pukapuka paturopi phage?

A.
E ai ki te pūtake o te whare pukapuka paturopi, ka taea te whakarōpū i ngā whare pukapuka paturopi phage ki roto i ngā whare pukapuka paturopi phage ārai mate me ngā whare pukapuka paturopi phage taketake. Ko te rerekētanga matua i waenga i ēnei momo e rua kei roto i te mea kua werohia ngā kararehe. Hei tāpiri, ka taea te whakaputa i ngā whare pukapuka paturopi taketake mā te hanga-haurua, te hanga-ā-ringa rānei.
P.
Me pēhea te aromatawai i te kounga o tētahi whare pukapuka paturopi phage?

A.
E rua ngā tohu matua mō te aromatawai i te kounga o tētahi whare pukapuka paturopi: te kaha rokiroki me te kanorau. Mā te kaha rokiroki nui, me te kanorau nui, ka whakarite kia whai hua te tirotiro i ngā paturopi me te kaha honohono me te motuhaketanga teitei.

Ngā Pātai Tirotiro Whare Pukapuka ◢

P.
He aha te tukanga mō te tirotiro i tētahi whare pukapuka paturopi phage?

A.
I roto i te tukanga whakakorenga koiora, ka whakatōkia te whare pukapuka whakaaturanga ki ngā ūnga pumau, kātahi ka horoia kia nui hei tango i ngā phage kore-tauhohe. Ko te tikanga ka whakakorea ngā hononga mā te whakamahi i tētahi otinga waikawa, tote nui rānei, ā, muri iho ka whakanuia ki roto i ngā pūtau manaaki e tika ana hei whakarei ake. I muri i te 3 ki te 5 rauna o te tirotiro, ka whiwhihia he paturopi he kaha te hononga.
P.
He aha ngā tikanga e whakamahia ana hei tirotiro i ngā whare pukapuka paturopi phage?

A.
Ko ngā tikanga tirotiro phage ko te tirotiro ā-pūtau totoka, te tirotiro ā-pūtau wai, me te tirotiro ā-pūtau. Me whai whakaaro te kōwhiringa o tētahi tikanga tirotiro tika ki ngā āhuatanga o ngā ngota whainga, ngā whāinga o te tukanga tirotiro, me te taiao taiwhanga.
P.
He aha ngā whakamātautau pai me ngā whakamātautau kino o ngā phage?

A.
Ko te tātari pai te tikanga tino taketake mō te tātari i ngā paturopi phage e herea ana ki ngā pūmua ūnga. Ko te tātari kino he whakakore i ngā paturopi kore-motuhake. I roto i te tukanga tātari paturopi phage, ka taea te tautuhi i ngā paturopi kore-motuhake nā te pokanoa mai i te tapanga antigen ūnga, ngā rauemi, ētahi atu antigen rānei he rite te hanganga ki te antigen ūnga. I roto i ēnei āhuatanga, ka taea e te tātari kino te whakarei ake i te whai huatanga o te tukanga tātari.
P.
Me pēhea te karo i te tirotiro i ngā paturopi kore-motuhake?

A.

Ka taea te whakarite i ngā paturopi kua tohua ki ngā momo tohu rerekē mō te tātaritanga tuatahi, ka taea rānei te whakamahi i te tātaritanga kino hei whakaiti i te ōwehenga o ngā paturopi kore-motuhake.

Ka taea e koe te whakakapi i te wai hiri e tika ana, te whakamahi rānei i ngā wai hiri rerekē hei ārai i tēnei āhuatanga. Mēnā ka tirotirohia te paturopi e mōhio ana ki te rārangi pūtau papamuri (CHO, HEK293 rānei), ka taea e koe te whakakapi i ngā rārangi pūtau papamuri rerekē mō ngā kaupapa tirotiro, tirotiro kino rānei.
P.
Me pēhea te manatoko me te tātari i ngā paturopi i whiwhihia?

A.
Ko ngā tikanga mō te manatoko i te paturopi ko te ELISA, te Western Blotting (WB), te flow cytometry, te immunoprecipitation (IP), te immunofluorescence (IF), te immunohistochemistry, me ētahi atu tikanga whakamātautau. He nui te wheako o Alpha Lifetech ki te kimi paturopi, ā, he rōpū whakamātautau ngaio anō hoki tā rātou, hei whakarite i te whai huatanga o ngā paturopi i ngā whakamātautau e whai ake nei.

Mena he pātai āu, tēnā whakapā mai ki a mātou i ngā wā katoa.

Leave Your Message

Ratonga Matua

Pūhara Kimi Paturopi