Tšebeletso ea Ntlafatso ea Li-antibody tsa Monoclonal

Alpha Lifetech Inc.e ka fana ka ponelopele ea epitope, tlhahiso ea peptide, tlhahiso ea protheine, tlhahiso ea li-antibody tsa polyclonal, kapa monoclonal hammoho le tlhoekiso ea li-antibody ho tsoa setsong sa sele, mokelikeli oa ascites kapa serum eohle.

Alpha Lifetech Inc.e na le boiphihlelo ba ho fa bareki lits'ebeletso tse fapaneng tsa mohala oa sele o tsitsitseng ho kenyeletsoa lisele tsa hybridoma, mela ea sele ea ho hlophisa genome, mela ea sele ea ho theola, le mela ea sele ea ho eketsa tlhahiso. Re leka ho fana ka lits'ebeletso tse tsoetseng pele tsa mohala oa sele ho moreki e mong le e mong 'me lits'ebeletso tsa rona li pakiloe ebile lia tšeptjoa.

Leano la Ntlafatso ea Li-antibody

1. Moralo le tokisetso ea antigen
Li-antigen tsena e ka ba liprotheine, li-peptide, kapa limolek'hule tse ling. Ho rala li-immunogen ho hloka mokhoa o akaretsang o kopanyang tsebo ea sebopeho sa antigen, thuto ea li-immunology le litsamaiso tsa ho pepa. Alpha Lifetech e na le theknoloji ea moralo oa li-antigen e ka ralang li-immunogen bakeng sa bareki ba hlokang.
Alpha Lifetech e fana ka dihlahiswa tsa protheine e kopantsweng hape bakeng sa bareki bao ba ka kgethang ho tsona. Re na le mefuta e fapaneng ya ditsamaiso tsa tlhahiso ya protheine, tse kang sistimi ya tlhahiso ya E. coli, sistimi ya tlhahiso ya tomoso, sistimi ya tlhahiso ya dikokonyana tsa baculovirus, sistimi ya tlhahiso ya diphoofolo tse anyesang, jj., ho hlahisa diprotheine tse kopantsweng hape tseo o di hlokang bakeng sa dipatlisiso.

2. Ho entoa liphoofolo
Alpha Lifetech e ka fana ka li-immunogen ho bareki, 'me liphoofolo tse itšireletsang mafung tse kang litoeba, mebutlanyana, linku, li-alpaca, likolobe tsa Guinea, li-hamster, jj. lia fumaneha hore u ka khetha ho tsona.

3. Mekhoa ea tlhahiso ea antibody ea monoclonal

●Tlhahiso ea Mabs ka theknoloji ea Hybridoma
Hybridoma ke sele e tsoakiloeng e hlahisoang ke ho kopanngoa ha mefuta e 'meli ea lisele, e leng mouse myeloma cell line le plasma cells (lymphocyte B), moo heterozygous cell e ka hlahisang li-antibodies tse tsoelang pele khahlanong le antigen e khahlisang in vitro. Tlhahiso ea hybridoma cell line e na le likarolo tse tharo: moralo oa antigen (haptens, limolek'hule tse nyane, le li-peptide), ente ea liphoofolo, ho kopanngoa ha lisele, le tlhahlobo ea clone e ntle. Ha re sebelisa mekhoa e ikhethang ea ente, re fumana bokhoni bo phahameng ka ho fetisisa ba ho kopanya lisele (li-hybridoma tse 1-10 ka lisele tse sekete tsa B) le li-antibody titers tse phahameng, ho eketsa sekhahla sa rona sa katleho litekong tse latelang tsa tlhahlobo tse kenyeletsang tlhahlobo ea biochemical, liteko tse thehiloeng liseleng tsa in vitro, le lithuto tsa liphoofolo.

●Tlhahiso ea Mabs ka theknoloji ea ponts'o ea phage
Pontšo ea phage e fana ka mokhoa o mong oa ho hlahisa li-antibodies tsa monoclonal. Li-B-lymphocyte li aroloa maling a liphoofolo 'me mRNA ea tsona ea ntšoa. Ka PCR, mRNA ena e fetoloa cDNA, e eketsang likarolo tsohle tsa VH le VL. Likarolo tsena ebe li kopanngoa ho ba vector, hangata e le likaroloana tse feto-fetohang tsa ketane e le 'ngoe (scFv), hammoho le protheine ea PIII ea bacteriophage. Ka mor'a moo, sebopeho sena se sebelisoa ho tšoaetsa E. coli, e leng se fellang ka ho thehoa ha laeborari e nang le lisele tse ka bang 10^10 ka ho enta ka phage e thusang. E. coli e khona ho ntša bacteriophage e nang le likarolo tsa VH le VL e le karolo ea seaparo sa eona. Ka mor'a sena, likarolo tse itseng tsa VH le VL tse shebaneng le antigen e khahlisang li ka khethoa 'me tsa sebelisoa ho tsosolosa E. coli ka bacteriophage. Lisele tse nang le plasmid li aroloa le ho aroloa ka tatellano.

4. Tlhahlobo ea motsoako

●Ho Kopanngoa ha Lisele le Khetho
Lisele tsa B tse kotutsoeng li kopanngoa le lisele tsa myeloma, e leng lisele tse nang le mofetše tse ka atang ka ho sa feleng ka har'a setso. Ho kopanngoa hangata ho finyelloa ka mokhoa o kang ho kopanya polyethylene glycol (PEG).
Kamora ho kopanngoa, lisele tsa hybridoma li lengoa ka mokhoa o ikhethileng o lumellang li-hybridoma feela ho phela. Hangata mokhoa ona o na le aminopterin, e thibelang kholo ea lisele tsa myeloma tse sa tsoakoang.

●Ho Hlahlobisisa le ho Kopanya
Lisele tse hlahang tsa hybridoma li hlahlojoa bakeng sa tlhahiso ea li-antibodies tse ikhethileng ho antigen e reriloeng. Sena hangata se etsoa ho sebelisoa mekhoa e kang enzyme-linked immunosorbent assay (ELISA), Western blot, immunoprecipitation, le flow cytometry.

Hang ha lisele tsa hybridoma tse hlahisang antibody li se li tsejoa, lia kopanngoa ho hlahisa palo ea lisele tse tšoanang. Sena se tiisa hore antibody e hlahisoang ke sele e 'ngoe le e 'ngoe ea hybridoma e tšoana.

5. Netefatso ea Ts'ebetso ea Antibody
Alpha Lifetech e fana ka netefatso ea ts'ebetso ea li-antibody, joalo ka Western blotting, immunohistochemistry, kapa liteko tsa ts'ebetso, ho khetholla li-antibody tsa monoclonal le ho netefatsa ho hlaka ha li-antibody, kamano le ts'ebetso.

6. Ntlafatso ea li-antibody
Ho hlwekisa di-antibodies ho tswa ho supernatant ya setso ho sebediswa mekgwa e kang protheine A kapa protheine G affinity chromatography, le Alpha Lifetech le tsona di ka fana ka mekgwa ya ho fetola di-antibody ho ntlafatsa affinity ya di-antibody.

7. Tlhahiso ea li-antibody
Alpha Lifetech e ka lekola li-antibodies tse ka khethoang bakeng sa lits'ebetso tsa tlhahlobo ea phallo e phahameng le tlhahiso e kholo.