Intangiriro ya serivisi yo kwerekana imiterere ya phage

Kugaragaza phage, nk'uburyo bwa molekile bushingiye ku guhindura ADN ya phage, byabaye igikoresho gikomeye cya molekile mu gukora isuzuma rya molekile ku ntego runaka binyuze mu kwerekana utwo duce ku buso bwa phage bityo bigatoranya uduce twa peptide/antibody dufite imiterere yihariye yo gufatana mu buryo butandukanye (amasomero).

Alpha Lifetech ifite uburambe bw'imyaka icumi mu guteza imbere uturemangingo tw’umubiri dukoresha ikoranabuhanga rya phage display antibodies kandi ishobora gutanga serivisi z’umwuga zo kubaka no gusuzuma isomero ry’umubiri ku bakiriya bacu. Dukurikije ibyo abakiriya bakeneye, dushobora gutanga ubwoko butandukanye bw’amasomero y’umubiri, nk’amasomero y’ubudahangarwa bw’umubiri, amasomero gakondo, amasomero y’ubukorano n’amasomero y’umubiri akoreshwa mu buryo bwa semi-synthetic. Ikoranabuhanga rya Phage display isomero, rihuza genes z’umubiri n’uturemangingo twa poroteyine za phage coat nka M13, ni ryo koranabuhanga rikunze gukoreshwa mu gutegura uturemangingo tw’umubiri kandi rikoreshwa cyane mu guteza imbere uturemangingo tw’umubiri twihariye cyane.

Alpha Lifetech izakingira alpaca, llama, alpaca, n'ifi zo mu bwoko bwa shark, n'ibindi, ku buryo imikorere yazo igera kuri 10^5. Tuzohereza raporo y'isuzuma ry'imikorere ku mukiriya kugira ngo tumenye neza ko amakuru ari ukuri kandi ari ay'ukuri. Amasomero gakondo afite ibyiza byo kuba afite uburozi buke kandi nta bushobozi bwo kurwanya indwara. Alpha Lifetech ifite umubare munini w'amasomero y'abasilikare kare yakorewe mbere, ashobora gukoreshwa mu buryo butaziguye mu gusuzuma abasilikare badakingiye inyamaswa, bityo bikagabanya igihe cy'umushinga, bigateza imbere iterambere ry'igerageza ry'umukiriya, kandi bikanizeza ko antibody ikora neza.

Igishushanyo cya 1: Ihame ryo kwerekana imiterere y'urukiramende

Sisitemu yacu yo kwerekana phage

Amakuru y'ingenzi ya sisitemu yo kwerekana M13/ T4/ T7 phage agaragara mu mbonerahamwe ya 1.

Imbonerahamwe ya 1 Ibinyabiziga bitandukanye byo kwerekana imiyoboro ya phage n'imiterere yabyo
	M13	T4	T7
Ingano ya genome	6407 bp	168895 bp	39937 bp
Poroteyine yo kwerekana	pVI, pIII na pVIII	SOC na HOC	gp10B
Ingano y'iyerekana	>110 kDa ni pIII
Ingano y'iyerekana
Ubucucike bw'ishusho
Ubucucike bw'ishusho
Inzira y'ubuzima	Lysogeny	Lytike	Lytike

Alpha Lfetech Ishobora Gutanga

Ishusho ya Phage ihuza ubwoko bwa genotype na phenotype mu buryo bumwe, ihuza guhitamo no kongera ubushobozi hamwe n'ubushobozi bukomeye bwo gusuzuma. Alpha Lifetech ishobora gutanga serivisi zitandukanye zo guteza imbere antibody ya phage display.
Serivisi z'ingenzi zirimo: Urubuga rw'isomero ry'abarwanyi ba VHH, urubuga rw'isomero ry'abarwanyi ba scFv, urubuga rw'isomero ry'abarwanyi ba Fab, urubuga rwo kubaka isomero rya phage, urubuga rwo gusuzuma isomero rya phage na serivisi yo gupima abarwanyi n'izindi serivisi.

iterambere ry'uturemangingo tw'umubiri - Alpha Lifetech

Igishushanyo cya 2: Uburyo bwo guteza imbere serivisi yo kwerekana imiterere y'urukiramende

Uburyo bwo guteza imbere ubudahangarwa bw'umubiri mu kwerekana phage

Dushobora gukingira inyamaswa dukoresheje poroteyine ziboneka muri laboratwari yacu cyangwa zitangwa n'abakiriya bacu, harimo poroteyine zitandukanye za macromolecular, peptides na poroteyine za membrane. Binyuze mu isuzuma rya ELISA, abashakashatsi bacu bakoze isesengura rya PBMC no gukuramo RNA mu maraso y'inkeri, maze batanga ibisubizo byiza by'ikingira. Dukoresheje uburyo bwo guhindura no kongera gene, twubatse gene kuri pMESC, pComb3XSS cyangwa pCANTAB 5E phage vectors. Dukoresheje electrotransformation y'uturemangingo dufite ubushobozi bwa TG1 E. coli, amasomero ya antibody ya phage afite ubushobozi bwo gusoma burenga 10^9 araboneka. Amasomero ya antibody afite ubushobozi bwo gusoma afite ubushobozi bwinshi bwo gusoma afite ubwoko butandukanye kandi afasha kubona antibody zifite ubushobozi bwo gusoma. Kugira ngo turebe ko isomero rya antibody rifite ubuziranenge, tuzagaragaza ubushobozi bwo gusoma no kwandika isomero. Amaherezo dukurikirana amasomero kugira ngo duhe abakiriya bacu sequences 30-50 zikenewe. Hanyuma, dukora isuzuma rya phage display Library, Ubwa mbere, antigen irahagarara kuri microplates za polypropylene, hanyuma binyuze mu isuzuma rya 3-5, phage-antibodies zifite ubushobozi bwo gufatana bugabanuka, maze clones zihariye zifatana na antigen zikagumaho. Ku isuzuma rya phage, uburyo bwa ELISA bukoreshwa kugira ngo buhereho clone nziza, mu gihe ku isuzuma ry'umusemburo, uburyo bwa FACS bukoreshwa kugira ngo uturemangingo tugaragaze uduce twa antibody dukoresheje antigens twanditseho. Binyuze mu isuzuma, dutegura antibody zifite ubushobozi bwo kumva, umwihariko no gukundana cyane.

Igishushanyo cya 3. Uburyo bwo gukora antibody ya Phage display

IBYIZA BYA SERIVISI
Ibyiza bya serivisi

Alpha Lifetech yiyemeje cyane ikoranabuhanga ryo kwerekana phage, kandi ishobora guha abakiriya serivisi zagutse kandi nziza zo kwerekana phage.

Igihe n'imikorere myiza cyane

Isosiyete yacu ishobora gushakisha antibody zihagije mu gihe gito; dushobora gukora igerageza ritandukanye ryo gusuzuma antibody hakurikijwe ibyo abakiriya bakeneye.

Ubwiza bw'ibicuruzwa buri hejuru

Isosiyete yacu ifasha abakiriya gutegura antibodies zifite umwihariko wo hejuru kandi zihamye, bikongera umusaruro w'ibyavuye mu bushakashatsi bwa siyansi.

Tanga amahitamo atandukanye

Isosiyete yacu ishobora kubaka amasomero y’abarwanyi b’ubwoko bwinshi (abantu, imbeba, urukwavu, alpaca, nibindi) kandi ishobora kubaka amoko atandukanye y’amasomero y’abarwanyi b’umubiri (scfv, Fab, VHH, nibindi).

Ubushobozi bunini

Isomero ryacu ry’abashinzwe kurwanya indwara rifite ubushobozi bwinshi bwo kurwanya indwara zirenze 10^9

Ibibazo Bikunze Kubazwa - Ibibazo Rusange byo Kugaragaza Phage

Ibibazo byo kwerekana phage ◢

Q.
Ikoranabuhanga ryo kwerekana phage ni iki?

A.
Ikoranabuhanga ryo kwerekana Phage ni ukwinjiza gene y’amahanga mu mwanya wihariye wa poroteyine ya phage capsid, no kugaragaza gene y’amahanga ku buso bwa phage nta ngaruka ku mikorere isanzwe ya phage. Nyuma y’isuzuma inshuro nyinshi, poroteyine y’ingenzi ifite umwihariko ukomeye n’imikorere myinshi ya biyolojiya yabonetse.
Q.
Ni izihe nyungu z'ikoranabuhanga ryo kwerekana phage?

A.
Amasomero y’abasilikare bafite ubushobozi bwinshi ashobora gukorwa hakoreshejwe ubu buryo. Igihe cyo gusuzuma ni gito, inzira yo gukora ni yoroshye, kandi uburyo bwo gukoresha ni bwinshi, bigatuma hamenyekana uburyo bwiza bwo gusuzuma peptides cyangwa poroteyine. Mu bikorwa bijyanye na virusi, abashakashatsi bashobora kuvumbura abasilikare bifatanya na epitopes zikomeye za virusi basuzuma amasomero manini y’abasilikare. Ubu buryo bushobora kubangamira uburyo virusi zinjira, zikora cyangwa zirinda ubudahangarwa bw’umubiri, bityo bigashyiraho urufatiro rwo kuvura indwara.
Q.
Ni irihe tandukaniro riri hagati y'ikoranabuhanga rya hybridoma n'ikoranabuhanga rya phage?

A.
Ikoranabuhanga ryo kwerekana udukoko duto (Phage display) rikoreshwa cyane kurusha ikoranabuhanga rya hybridoma, rigarukira gusa ku gukora udukoko duto duto turwanya ubudahangarwa bw’umubiri runaka. Mu buryo bunyuranye, ikoranabuhanga ryo kwerekana udukoko duto rishobora kwerekana isomero ryose ry’udukoko dukomoka ku bwoko butandukanye bw’ubudahangarwa bw’umubiri.
Q.
Ni izihe ngamba ikoranabuhanga ryo kwerekana phage rikoreshwa?

A.
Ikoranabuhanga ryo kwerekana udukoko duto (Phage ecran) rifite uburyo bwinshi butandukanye bwo gukoresha, harimo gusuzuma uduce twakira udukoko duto dutera indwara, kwiga uburyo poroteyine ikorana, gukurikirana uburyo ibimenyetso bikwirakwira hagati y’uturemangingo, gusuzuma indwara z’inyamaswa, no gukora inkingo, n’ibindi.
Q.
Ese hari icyizere cy'ikoranabuhanga ryo kwerekana phage?

A.
Ikoranabuhanga ryo kwerekana udukoko duto (Phage display) ritanga ibyiza byinshi kandi rishobora gukoreshwa mu nzego zitandukanye. Ariko, ikibazo cy'ingenzi mu guteza imbere antibodies recombinant hakoreshejwe uburyo bwo kwerekana udukoko duto (phage display) ni uburyo bugoye n'igiciro bijyana n'iyi gahunda. Ubu buryo busaba guhuza tekiniki za molekile, harimo guhindura imiterere y'uturemangingo, guhindura imiterere y'uturemangingo, no kugaragaza ibimenyetso, hamwe n'isesengura ry'ubudahangarwa bw'umubiri, ibinyabutabire, na biophysical kugira ngo hamenyekane kandi hamenyekane antibodies zifuzwa.

Ibibazo bifitanye isano n'imisemburo y'uturemangingo ◢

Q.
Ni ubuhe buryo bwo kwerekana antigen?

A.
Ukwigaragaza kwa antigen ni ingenzi cyane. Uburyo bwo kwerekana antigen burimo ahanini kwerekana antigen ikoze mu buryo butaziguye, kwerekana antigen ikozwe mu buryo butaziguye, kwerekana antigen binyuze mu gusuzuma uturemangingo twose, kwerekana antigen binyuze muri liposome, nano-phospholipid disks na VLPs.
Q.
Ni iyihe tandukaniro riri hagati y’ubwoko butandukanye bw’ikwirakwizwa rya antigen?

A.
Uburyo bwo gutwikira mu buryo butaziguye bworoshye, ariko imiterere ya antigen yoroshye guhindura, kandi imikorere yo gutwikira mu buryo butaziguye iragoye, ariko ishobora gukomeza imiterere ya antigen no kunoza imikorere myiza yo kwerekana. Gusuzuma uturemangingo bishobora kwigana aho ibintu bibera mu mubiri, ariko hari ikibazo cyo gufatana ku buryo budasanzwe. Disiki ya Nano-phospholipid ifite ingano nto, ikora neza cyane kandi ihamye neza; VLPs ihuzwa na antigen cyangwa ipfunyitse na antigen, ifite ubushobozi bwo kwirinda indwara nyinshi n'umutekano mwiza, kandi ishobora gutera ubudahangarwa bukomeye bw'umubiri.
Q.
Ni iyihe ngaruka y'ubwinshi bw'abarwayi ba antigen ku bisubizo by'ikoranabuhanga ryo kwerekana phage?

A.
Iyo ingano y’ibice iri hasi cyane, amahirwe yo gufatana na phage azagabanuka, bigatuma ubushobozi bwo gusuzuma bugabanuka. Ku rundi ruhande, ingano ikabije ishobora kongera amahirwe yo gufatana ku buryo butari bwihariye, ikazamura ikimenyetso cy’inyuma, kandi ikabangamira kumenya imiterere y’ibice byiza.
Q.
Ni gute twakwizeza ko antigen igumana imiterere yayo karemano mu gihe cyo gusuzuma?

A.
Uburyo bworoshye bwo gukosora bushobora gukoreshwa kugira ngo hirindwe ubushyuhe bwinshi, aside ikomeye na alkali. Uburyo bwo gutwikira butaziguye hamwe na sisitemu ya biotin-streptavidin byakoreshejwe kugira ngo bigabanye kwangirika kw'imiterere ya antigen. Niba antigen ari poroteyine y'urukiramende, ishobora kugaragara ku buso bw'uturemangingo twangiritse cyangwa igakoreshwa mu kwigana ibidukikije bya membrane, nka nano-disk, kugira ngo igumane imiterere ya poroteyine y'urukiramende karemano.
Q.
Ni gute wagenzura imiterere y’udukoko twapimwe hakoreshejwe ikoranabuhanga rya phage display kuri antigens?

A.
Uretse igerageza risanzwe nka ELISA, Western Blotting (WB), na Immunofluorescence (IF), ubuhanga bwo gupima Surface Plasmon Resonance (SPR) na Bio-Layer Interferometry (BLI) bushobora gukoreshwa mu kugenzura inzira zo gufatanya no gutandukanya antibody na antigens mu gihe nyacyo, bigatuma hamenyekana uburyo bwo gufatana no gutandukanya ibintu.

Ibibazo bifitanye isano n'ubudahangarwa bw'umubiri ◢

Q.
Ni izihe ngamba muri rusange z’ubwirinzi bw’umubiri bw’inyamaswa mu ikoranabuhanga ryo kwerekana phage?

A.
Ishusho ya phage ishobora gukoreshwa mu gukora antibodies za monoclonal zifite affinity nyinshi ku bwoko bwinshi bw’abarwanya, harimo ariko ntizigarukira ku bantu, imbeba, imbeba, inkwavu, inkoko, ingamiya, alpacas, inka, imbwa, intama, inkende, n’ifi.
Q.
Ni ibihe biranga kandi ni ibihe bintu bifatika by’amasomero y’abarwanyi bakomoka ku moko atandukanye y’inyamaswa mu ikoranabuhanga ryo kwerekana phage?

A.
Ikoranabuhanga ryo kubaka amasomero y’abarwanyi b’imbeba rirazwi cyane, rirahendutse, kandi rimara igihe gito mu igerageza, bigatuma rikoreshwa mu gusuzuma abarwanyi mbere y’uko batangira ndetse no mu bushakashatsi bw’ibanze. Abarwanyi b’imbeba bagaragaza ubwiza n’umwihariko, bafite urwego rwagutse rwo kumenya epitope ugereranije n’abarwanyi b’imbeba. Abarwanyi b’imbeba bashobora kuboneka mu masomero y’abarwanyi b’imbeba, bityo birinda ingaruka zishobora kubaho iyo abarwanyi b’imbeba batanzwe mu mubiri w’umuntu. Ubu buryo bufite akamaro kanini mu bushakashatsi no mu iterambere ry’imiti, cyane cyane mu gukora abarwanyi b’imbeba bavura.
Q.
Ni iyihe immunogen rusange y'inyamaswa ikoreshwa mu ikoranabuhanga ryo kwerekana phage?

A.
Ubudahangarwa bw’umubiri bw’inyamaswa muri rusange bugabanyijemo poroteyine antigen, polypeptide antigen, nucleic acid antigen na antigen ifitanye isano n’indwara, muri zo antigen ifitanye isano n’indwara irimo antigen ya virusi (agace kuzuye ka virusi, poroteyine ya virusi, poroteyine idashingiye ku miterere ya virusi, nibindi), antigen ya bagiteri (igicuruzwa cyuzuye cya bagiteri, igice cy’urukuta rw’uturemangingo, poroteyine isohoka, uburozi, nibindi) na antigen y’udukoko.
Q.
Ni gute twakemura ikibazo cyo kwihanganira ubudahangarwa bw'umubiri mu nyamaswa mu gihe cy'ubwirinzi bw'umubiri?

A.
Hindura imiterere y’umubiri ukoresheje antigens z’ubudahangarwa bw’umubiri ukoresheje poroteyine zisubiramo hanyuma ushyireho uduce twa virusi zirwanya inkongi hamwe na poroteyine z’imiterere ya virusi. Hindura imiterere y’igerageza hanyuma uhindure ingano y’ubudahangarwa bw’umubiri.
Q.
Ni gute ubushobozi bw'ubudahangarwa bw'umubiri mu ikoranabuhanga ryo kwerekana phage bwakongerwa?

A.
Ubudahangarwa bw'umubiri bushobora kongerwa no guhuza antigens na poroteyine zitwara, nka albumin yo mu maraso y'inka. Byongeye kandi, ibikoresho by'ubudahangarwa bw'umubiri, harimo na Freund's adjuvant na aluminium adjuvant, bishobora gukoreshwa mu kongera igihe cyo kubaho mu mubiri no gushishikariza uturemangingo tw'ubudahangarwa gukora no kwiyongera, bityo bikanoza ubushobozi bw'ubudahangarwa bw'umubiri.

Ibibazo byo Kubaka Isomero ◢

Q.
Isomero ry'umubiri w'abarwanyi ba phage ni iki?

A.
Isomero ry’abarwanyi ba phage ni uruhurirane rw’abarwanyi ba phage batandukanye bakozwe no guteranya ubwoko bwose bw’uturemangingo tw’uturere dutandukanye duturutse kuri antibodies mu turemangingo twa phage hanyuma tukagaragara ku buso bwa phages. Ubu buryo butuma habaho ishingwa ry’abarwanyi ba phage.
Q.
Uburyo bwo kubaka isomero ry'umubiri urwanya indwara za phage

A.
Uburyo rusange bwo kwerekana phage bukubiyemo intambwe nyinshi z'ingenzi: gukuramo uturemangingo tw’amaraso two mu maraso (PBMC) mu nyamaswa zakingiwe, gutandukanya RNA yose no kuyihinduramo ADN yuzuzanya (cDNA), kongera gene y’ingenzi hakoreshejwe ikoranabuhanga rya polymerase chain reaction (PCR), gushyira gene y’ingenzi mu gukora phage vector, no guhindura vector mu turemangingo kugira ngo tugaragaze. Iyi gahunda ituma hakorwa isomero ry’igaragaza phage, rishobora kumenya vuba uturemangingo tw’umubiri dufite affinity. Nyuma y’ibyo, iri somero rishobora kuba isoko y’agaciro mu iterambere ry’inkingo, imiti ivura, n’ibikoresho byo gusuzuma.
Q.
Ni izihe ngingo z'ingenzi zo kubaka isomero ryiza?

A.

Menya neza ko umubare wose wa PBMC uhagije. Menya neza ko ubwiza bwa RNA butangiritse.

Kugira ngo umubare w’abahindura ibintu uboneke, ubusanzwe ubushobozi bw’isomero bugomba kuba inshuro 10 kugeza ku 100 z’ubusumbane bw’inyigisho.
Q.
Ni izihe nzego za 'phage antibody' zishyirwa mu byiciro?

A.
Dukurikije isoko y’isoko ry’ama-antibody, amasomero y’ama-antibody ya phage ashobora gushyirwa mu byiciro bibiri: amasomero y’ama-antibody ya phage na somero y’ama-antibody ya phage. Itandukaniro ry’ibanze hagati y’ubwo bwoko bubiri ni uko inyamaswa zakingiwe. Byongeye kandi, amasomero y’ama-antibody ya gakondo ashobora gukorwa binyuze mu gukora cyangwa gukora.
Q.
Ni gute umuntu yasuzuma ubwiza bw'isomero ry'umubiri w'abarwanyi ba phage?

A.
Hari ibipimo bibiri by'ingenzi byo gusuzuma ubwiza bw'isomero ry'abarwayi ba antibody: ubushobozi bwo kubika no gutandukanya. Ubushobozi bwinshi bwo kubika, hamwe n'ubwoko bwinshi, butuma habaho gusuzuma neza abarwayi ba antibody bafite ubwinshi n'uburyo bwihariye.

Ibibazo byo gusuzuma isomero ◢

Q.
Ni iyihe nzira yo gusuzuma isomero ry’umubiri w’abarwanyi ba phage?

A.
Mu gihe cyo gusesengura ibintu mu buryo bw'ibinyabuzima, isomero ry'aho ibintu bigaragarira rishyirwamo intego zihamye hanyuma rikagezwa cyane kugira ngo rikureho uduce tudakora neza. Uduce duto dukunze gukurwamo hakoreshejwe umuti wa aside cyangwa umunyu mwinshi hanyuma twongerwa mu tunyangingo twakira ibintu kugira ngo birusheho kuba byiza. Nyuma y'isuzuma ry'inshuro 3 kugeza kuri 5, haboneka utunyangingo dufite imbaraga nyinshi.
Q.
Ni ubuhe buryo bukoreshwa mu gusuzuma amasomero y’abarwanyi ba phage?

A.
Uburyo bwo gusuzuma phage burimo ahanini gupima ibice bikomeye, gusuzuma ibice by'amazi, no gusuzuma uturemangingo. Guhitamo uburyo bukwiye bwo gusuzuma bigomba kuzirikana imiterere y'uturemangingo twibandwaho, intego z'uburyo bwo gusuzuma, n'ibidukikije byo muri laboratwari.
Q.
Ni ibihe bipimo byiza n'ibibi bya phages?

A.
Gupima neza ni bwo buryo bw'ingenzi bwo gusuzuma abasirikari b'umubiri ba phage bafatana na poroteyine. Gupima nabi bifasha gukuraho abasirikari b'umubiri batari basanzwe. Mu gihe cyo gusuzuma abasirikari b'umubiri ba phage, abasirikari b'umubiri batari basanzwe bashobora kumenyekana bitewe n'ingaruka ziturutse ku kirango cya antigen, ibikoresho, cyangwa izindi antigen zisa n'iz'umubiri wa antigen. Muri ibyo bihe, gusuzuma nabi bishobora kongera imikorere myiza y'igikorwa cyo gusuzuma.
Q.
Ni gute wakwirinda gusuzuma antibodies zitari iz’umwihariko?

A.

Antigens zanditseho ubwoko butandukanye bw'ibimenyetso zishobora gushyirwa imbere mu gusuzuma bwa mbere, cyangwa gusuzuma ibitari byo bishobora gukoreshwa mu kugabanya umubare w'abasilikare badafite umwihariko.

Ushobora guhitamo gusimbuza amazi yo gufunga akwiye cyangwa ugakoresha andi mazi yo gufunga kugira ngo wirinde iki kibazo. Iyo antibody imenya umurongo w’uturemangingo tw’inyuma (CHO cyangwa HEK293) ipimwa, ushobora gusimbuza imirongo itandukanye y’uturemangingo tw’inyuma kugira ngo upime cyangwa upime ibitari byiza.
Q.
Ni gute umuntu yagenzura kandi agasesengura ubudahangarwa bw’umubiri bwabonetse?

A.
Uburyo bwo kugenzura ubudahangarwa bw’umubiri muri rusange burimo ELISA, Western Blotting (WB), flow cytometry, immunoprecipitation (IP), immunofluorescence (IF), immunohistochemistry, n'ubundi buryo butandukanye bwo kugerageza. Alpha Lifetech ifite uburambe bwinshi mu gupima ubudahangarwa bw’umubiri kandi ifite itsinda ry’inzobere mu kugerageza, rigenzura ko ubudahangarwa bw’umubiri bukora neza mu bushakashatsi bukurikira.

Niba ufite ikibazo icyo ari cyo cyose, twandikire igihe icyo ari cyo cyose.

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Urubuga rwo kuvumbura ubudahangarwa bw'umubiri