ʻO nā antibody, i kapa ʻia ʻo immunoglobulins (Ig), he glycoproteins e hoʻopaʻa pono i nā antigens.
 
Hana ʻia ka hoʻomākaukau antibody maʻamau e ka pale ʻana i nā holoholona a me ka hōʻiliʻili antiserum.No laila, loaʻa pinepine ka antiserum i nā antibodies e kūʻē i nā antigens pili ʻole a me nā ʻāpana protein ʻē aʻe i loko o ka serum.Loaʻa ka nui o nā molekele antigen maʻamau i nā epitopes like ʻole, no laila he hui pū ʻia nā antibodies maʻamau i nā epitopes like ʻole.ʻO nā antibodies serum maʻamau i kuhikuhi ʻia e kūʻē i ka epitope hoʻokahi i hoʻokumu ʻia i nā antibodies heterogeneous i hana ʻia e nā clones cell B like ʻole.No laila, kapa ʻia nā antibodies serum maʻamau i nā antibodies polyclonal, a i ʻole polyclonal antibodies no ka pōkole.
 
ʻO ka antibody monoclonal (monoclonal antibody) he antibody ʻano like ʻole i hana ʻia e kahi clone cell B hoʻokahi a kuhikuhi wale ʻia i kahi epitope kikoʻī.Hoʻomākaukau maʻamau ia e ka ʻenehana hybridoma—hoʻokumu ʻia ka ʻenehana antibody Hybridoma ma luna o ka ʻenehana fusion cell, e hui pū ana i nā cell B me ka hiki ke huna i nā antibodies kikoʻī a me nā cell myeloma me ka hiki ke ulu ʻole i loko o nā B-cell hybridomas.Loaʻa i kēia cell hybridoma nā ʻano o kahi cell makua.Hiki iā ia ke hoʻonui mau loa a make ʻole i loko o ka vitro e like me nā cell myeloma, a hiki iā ia ke synthesize a hūnā i nā antibodies kikoʻī e like me nā lymphocytes splenic.Ma o ka cloning, hiki ke kiʻi ʻia kahi laina monoclonal i loaʻa mai kahi cell hybridoma hoʻokahi, ʻo ia hoʻi, kahi laina kelepona hybridoma.ʻO nā antibodies i hana ʻia he mau antibodies homogenous kūʻē i ka mea hoʻoholo antigenic like, ʻo ia hoʻi, nā antibodies monoclonal.
 
Loaʻa nā mea ʻokoʻa ma ke ʻano he hoʻokahi a ʻoi aʻe paha nā monomers me ke ʻano Y (ʻo ia hoʻi, nā antibodies monoclonal a i ʻole polyclonal antibodies).Hoʻokumu ʻia kēlā me kēia monomer Y-like me 4 mau kaulahao polypeptide, me ʻelua kaulahao kaumaha like a me ʻelua kaulahao māmā like.Ua kapa ʻia ke kaulahao māmā a me ke kaulahao kaumaha e like me ko lākou kaumaha molekala.ʻO ka ʻaoʻao kiʻekiʻe o ka hoʻolālā Y-like ka ʻāpana ʻokoʻa, ʻo ia ka pae hoʻopaʻa antigen.(Mahele mai Detai Bio-Monoclonal Antibody Concept)
 
ʻO ka hoʻolālā antibody
kaulahao kaumaha
ʻElima ʻano o nā kaulahao kaumaha Ig mammalian, i kapa ʻia me nā hua Helene α, δ, ε, γ, a me μ.ʻO nā antibodies pili i kapa ʻia ʻo IgA, IgD, IgE, IgG, a me IgM.ʻOkoʻa nā kaulahao kaumaha like ʻole i ka nui a me ka haku mele ʻana.Aia i loko o ka α a me ka γ ma kahi o 450 amino acids, ʻoiai ʻo μ a me ε aia ma kahi o 550 amino acids.
Loaʻa i kēlā me kēia kaulahao kaumaha ʻelua mau ʻāpana: ka ʻāina mau a me ka ʻāpana hoʻololi.Loaʻa i nā antibodies o ka ʻano like ka ʻāpana mau, akā aia nā ʻokoʻa ma waena o nā antibodies o nā ʻano like ʻole.ʻO nā ʻāpana mau o nā kaulahao koʻikoʻi γ, α, a me δ ua hui pū ʻia me ʻekolu mau kikowaena Ig i hui pū ʻia, me kahi ʻāpana hinge e hoʻonui ai i kona maʻalahi;ʻo nā ʻāpana mau o nā kaulahao kaumaha μ a me ε i haku ʻia me 4 Ig domains.ʻOkoʻa ka ʻāpana ʻokoʻa o ke kaulahao kaumaha o ka antibody i hana ʻia e nā ʻāpana B like ʻole, akā ʻokoʻa ka ʻāpana ʻokoʻa o ka antibody i hana ʻia e ka cell B like a i ʻole cell clone, a ʻo ka ʻāpana ʻāpana o kēlā me kēia kaulahao kaumaha ma kahi o 110 amino acid ka lōʻihi., A hana i hoʻokahi domain Ig.
 
kaulahao māmā
ʻElua wale nō ʻano o nā kaulahao māmā i nā mammals: ʻano lambda a me ke ʻano kappa.Loaʻa i kēlā me kēia kaulahao māmā nā kāʻei kapu ʻelua: kahi ʻāina mau a me kahi ʻāpana ʻokoʻa.ʻO ka lōʻihi o ke kaulahao māmā e pili ana i 211 ~ 217 amino acids.ʻO nā kaulahao māmā ʻelua i loko o kēlā me kēia antibody ua like mau.No nā mammals, hoʻokahi wale nō ʻano o ke kaulahao māmā i kēlā me kēia antibody: kappa a i ʻole lambda.I loko o kekahi mau vertebrates haʻahaʻa, e like me nā iʻa cartilaginous (nā iʻa cartilage) a me nā iʻa iwi, ʻike pū ʻia nā ʻano kaulahao māmā ʻē aʻe e like me ke ʻano iota (iota).
 
Nā ʻāpana Fab a me Fc
Hiki ke hoʻohui pololei ʻia ka māhele Fc me nā enzymes a i ʻole nā ​​mea hoʻonaninani fluorescent e hōʻailona i nā antibodies.ʻO ia ka ʻāpana kahi i hoʻopaʻa ʻia ai ka antibody ma ka pā i ka wā o ka hana ELISA, a ʻo ia hoʻi ka ʻāpana kahi i ʻike ʻia a hoʻopaʻa ʻia ka antibody ʻelua i immunoprecipitation, immunoblotting a me immunohistochemistry.Hiki ke hoʻoheheʻe ʻia nā antibodies i ʻelua ʻāpana F(ab) a me hoʻokahi ʻāpana Fc e nā enzyme proteolytic e like me ka papain, a i ʻole hiki ke wāwahi ʻia mai ka ʻāpana hinge e ka pepsin a hoʻoheheʻe ʻia i hoʻokahi ʻāpana F(ab)2 a me hoʻokahi māhele Fc.He mea pono loa nā ʻāpana antibody IgG i kekahi manawa.Ma muli o ka nele o ka māhele Fc, ʻaʻole e kahe ka māhele F(ab) me ka antigen, ʻaʻole hoʻi e hopu ʻia e nā cell immune i loko o nā haʻawina vivo.Ma muli o nā ʻāpana liʻiliʻi liʻiliʻi a me ka nele o ka hana cross-linking (ma muli o ka nele o ka māhele Fc), hoʻohana pinepine ʻia ka ʻāpana Fab no ka radiolabeling i nā haʻawina hana, a ʻo ka māhele Fc ka mea i hoʻohana nui ʻia ma ke ʻano he mea pale i ka histochemical staining.
 
Nā ʻāpana like ʻole a mau
Aia ka ʻāpana hoʻololi ('āpana V) ma 1/5 a i ʻole 1/4 (ma kahi o 118 mau koena amino acid) o ke kaulahao H ma kahi kokoke i ka N-terminus a me 1/2 (ma kahi o 108-111 mau koena amino acid) kokoke i ka N-terminus o ke kaulahao L.Loaʻa i kēlā me kēia māhele V kahi apo peptide i hoʻokumu ʻia e nā kaulahao disulfide intra-chain, a aia kēlā me kēia apo peptide ma kahi o 67 a 75 mau koena amino acid.ʻO ka hui ʻana a me ka hoʻonohonoho ʻana o nā waikawa amino ma ka ʻāpana V e hoʻoholo i ka kikoʻī paʻa antigen o ka antibody.Ma muli o nā ʻano loli mau a me ke kaʻina o nā waikawa amino ma ka ʻāina V, hiki ke hana ʻia nā ʻano antibodies me nā ʻano kikoʻī antigen like ʻole.ʻO nā ʻāpana V o ke kaulahao L a me ke kaulahao H ua kapa ʻia ʻo VL a me VH.Ma VL a me VH, ʻoi aku ka kiʻekiʻe o ka hoʻololi ʻana o ka ʻakika amino a me ke kaʻina o kekahi mau wahi kūloko.Ua kapa ʻia kēia mau ʻāina he hypervariable regions (HVR).ʻO ka hui ʻana o ka waikawa amino a me ka hoʻonohonoho ʻana o nā ʻāpana non-HVR ma ka ʻāpana V he ʻano conservative, i kapa ʻia ʻo ka ʻāpana framework.Aia ʻekolu mau ʻāpana hypervariable ma VL, aia ma nā koena waikawa amino 24 a 34 a me 89 a 97.ʻO nā HVR ʻekolu o VL a me VH ua kapa ʻia ʻo HVR1, HVR2 a me HVR3.ʻO ka noiʻi a me ka nānā ʻana o ka ʻokoʻa aniani X-ray i hōʻike ʻia ʻo ka ʻāina hypervariable ka wahi i hoʻopaʻa ʻia ai ka antigen antibody, no laila ua kapa ʻia ʻo ia ka complementarity-determining region (CDR).Hiki ke kapa ʻia ka HVR1, HVR2 a me HVR3 o VL a me VH CDR1, CDR2 a me CDR3.ʻO ka maʻamau, ʻoi aku ka kiʻekiʻe o ka CDR3 o ka hypervariability.ʻO ka ʻāpana hypervariable ka wahi nui kahi e noho ai nā mea hoʻoholo idiotypic o nā molekala Ig.I ka hapanui o nā hihia, ʻoi aku ka nui o ke kaulahao H i ka hoʻopaʻa ʻana i ka antigen.
ʻO ka ʻāpana mau (C region)Aia ma 3/4 a i ʻole 4/5 (ma kahi o ka amino acid 119 a i ka C terminal) o ke kaulahao H kokoke i ka C terminal a me 1/2 (loaʻa ma kahi o 105 amino acid koena) kokoke i ka C terminal o ke kaulahao L.Loaʻa i kēlā me kēia māhele hana o ke kaulahao H ma kahi o 110 mau koena amino acid, a he apo peptide i haku ʻia me 50-60 mau koena amino acid i hoʻopili ʻia e nā paʻa disulfide.ʻO ka hui ʻana o ka waikawa amino a me ka hoʻonohonoho ʻana o kēia ʻāina he mau maʻamau i loko o ka holoholona like Ig isotype L kaulahao a me ke kaulahao H ʻano like.ʻO ka mea like, hiki iā ia ke hoʻopaʻa paʻa wale i ka antigen e pili ana, akā ʻo ke ʻano o kāna ʻāpana C he like, ʻo ia hoʻi, he like ka antigenicity.Hiki ke hoʻohui ʻia ka lio anti-human IgG secondary antibody (a i ʻole anti-antibody) me ʻelua A hui pū ʻia o nā antibodies (IgG) e kūʻē i nā exotoxins like ʻole.He kumu koʻikoʻi kēia no ka hoʻomākaukau ʻana i nā antibodies lua a me ka hoʻohana ʻana i ka fluorescein, isotopes, enzymes a me nā mea ʻē aʻe i hōʻailona ʻia.
 
 
Nā huahana pili:
Kelepona pololei RT-qPCR kime