Ma antibodies, omwe amatchedwanso ma immunoglobulins (Ig), ndi ma glycoprotein omwe amamangiriza ku ma antigen.
 
Kukonzekera kokhazikika kwa ma antibodies kumapangidwa ndi katemera wa nyama ndikusonkhanitsa antiserum.Choncho, antiserum nthawi zambiri imakhala ndi ma antibodies motsutsana ndi ma antigen ena osagwirizana ndi zigawo zina za mapuloteni mu seramu.Ma antigen mamolekyu ambiri amakhala ndi ma epitopes osiyanasiyana, kotero kuti ma antibodies wamba amakhalanso osakanikirana a ma antibodies motsutsana ndi ma epitopes osiyanasiyana.Ngakhale ma antibodies wamba a seramu omwe amalimbana ndi ma epitope omwewo amakhalabe ndi ma antibodies osiyanasiyana opangidwa ndi mitundu yosiyanasiyana ya ma cell a B.Chifukwa chake, ma antibodies wamba a seramu amatchedwanso ma antibodies a polyclonal, kapena ma antibodies a polyclonal mwachidule.
 
Antibody ya Monoclonal (monoclonal antibody) ndi antibody yofanana kwambiri yopangidwa ndi gulu limodzi la B cell ndipo imangolunjika pa epitope inayake.Nthawi zambiri amakonzedwa ndi teknoloji ya hybridoma-teknoloji ya hybridoma antibody imachokera ku teknoloji yophatikizira ma cell, kuphatikiza ma cell a B omwe amatha kutulutsa ma antibodies enieni ndi ma cell a myeloma omwe ali ndi mphamvu yakukula kosalekeza kukhala B-cell hybridomas.Selo la hybridoma ili ndi mawonekedwe a selo la kholo.Imatha kuchulukirachulukira kosatha komanso kosatha mu vitro ngati maselo a myeloma, ndipo imatha kupanga ndikutulutsa ma antibodies enieni ngati ma splenic lymphocytes.Kupyolera mu cloning, mzere wa monoclonal wochokera ku selo limodzi la hybridoma, ndiko kuti, mzere wa hybridoma cell, ukhoza kupezeka.Ma antibodies omwe amapanga ndi ma antibodies ofanana kwambiri ndi antigenic determinant, ndiko kuti, ma antibodies a monoclonal.
 
Ma antibodies amakhala ngati ma monomers amodzi kapena angapo ooneka ngati Y (mwachitsanzo, ma antibodies a monoclonal kapena ma polyclonal antibodies).Monoma iliyonse yooneka ngati Y imapangidwa ndi maunyolo 4 a polypeptide, kuphatikiza maunyolo awiri olemera ofanana ndi maunyolo awiri ofanana.Unyolo wopepuka ndi unyolo wolemera amatchulidwa molingana ndi kulemera kwawo kwa maselo.Pamwamba pa mawonekedwe a Y ndi malo osinthika, omwe ndi malo omangira antigen.(Katundu wa Detai Bio-Monoclonal Antibody Concept)
 
Kapangidwe ka ma antibodies
Unyolo wolemera
Pali mitundu isanu ya maunyolo olemera a mammalian Ig, otchulidwa ndi zilembo zachi Greek α, δ, ε, γ, ndi μ.Ma antibodies ofanana nawo amatchedwa IgA, IgD, IgE, IgG, ndi IgM.Unyolo wolemera wosiyanasiyana umasiyana kukula ndi kapangidwe.α ndi γ zili ndi pafupifupi 450 amino acid, pamene μ ndi ε zili ndi pafupifupi 550 amino acid.
Unyolo uliwonse wolemera uli ndi zigawo ziwiri: chigawo chokhazikika ndi dera losinthika.Ma antibodies onse amtundu umodzi amakhala ndi dera lofanana, koma pali kusiyana pakati pa ma antibodies amitundu yosiyanasiyana.Madera okhazikika a maunyolo olemera γ, α, ndi δ amapangidwa ndi madera atatu a Ig motsatizana, ndi dera la hinge kuti awonjezere kusinthasintha kwake;madera okhazikika a unyolo wolemera μ ndi ε amapangidwa ndi madera a 4 Ig.Chigawo chosinthika cha unyolo wolemera wa ma antibody opangidwa ndi ma B ma cell osiyanasiyana ndi osiyana, koma dera losinthika la antibody lomwe limapangidwa ndi B cell lomwelo kapena cell clone ndilofanana, ndipo gawo losinthika la unyolo uliwonse wolemera ndi pafupifupi 110 amino acid m'litali., Ndipo pangani dera limodzi la Ig.
 
Unyolo wopepuka
Pali mitundu iwiri yokha ya maunyolo owala pa nyama zoyamwitsa: mtundu wa lambda ndi mtundu wa kappa.Unyolo uliwonse wowunikira uli ndi madera awiri olumikizidwa: chigawo chokhazikika ndi dera losinthika.Kutalika kwa unyolo wowala ndi pafupifupi 211 ~ 217 amino acid.Maunyolo awiri owala omwe ali mu antibody iliyonse amakhala ofanana nthawi zonse.Kwa zoyamwitsa, tcheni chowunikira mu antibody iliyonse chimakhala ndi mtundu umodzi wokha: kappa kapena lambda.M'zamoyo zina zam'munsi, monga nsomba za cartilage (cartilage fishes) ndi bony fish, mitundu ina ya maunyolo opepuka monga mtundu wa iota (iota) amapezekanso.
 
Fab ndi Fc magawo
Gawo la Fc litha kuphatikizidwa mwachindunji ndi ma enzymes kapena utoto wa fulorosenti kuti ulembe ma antibodies.Ndi gawo lomwe antibody rivets pa mbale pa ndondomeko ya ELISA, komanso ndi gawo limene antibody yachiwiri imadziwika ndikumangidwa mu immunoprecipitation, immunoblotting ndi immunohistochemistry.Ma antibodies amatha kupangidwa ndi hydrolyzed kukhala magawo awiri a F(ab) ndi gawo limodzi la Fc ndi ma enzymes a proteolytic monga papain, kapena amatha kusweka kuchokera ku hinge dera ndi pepsin ndikupangidwa ndi hydrolyzed kukhala gawo limodzi la F(ab)2 ndi gawo limodzi la Fc.Zidutswa za antibody za IgG nthawi zina zimakhala zothandiza kwambiri.Chifukwa cha kusowa kwa gawo la Fc, gawo la F (ab) silidzathamanga ndi antigen, komanso silidzagwidwa ndi maselo a chitetezo cha mthupi mu maphunziro a vivo.Chifukwa cha tizidutswa tating'ono tating'onoting'ono ta maselo ndi kusowa kwa ntchito yolumikizirana (chifukwa cha kusowa kwa gawo la Fc), gawo la Fab nthawi zambiri limagwiritsidwa ntchito popanga ma radiolabeling mu maphunziro ogwirira ntchito, ndipo gawo la Fc limagwiritsidwa ntchito makamaka ngati chotchinga mu histochemical staining.
 
Madera osinthika komanso osasintha
The variable dera (V dera) ili pa 1/5 kapena 1/4 (yokhala pafupifupi 118 amino asidi zotsalira) wa H unyolo pafupi N-terminus ndi 1/2 (wokhala za 108-111 amino asidi zotsalira) pafupi N-terminus wa L unyolo.Chigawo chilichonse cha V chimakhala ndi mphete ya peptide yopangidwa ndi ma intra-chain disulfide bond, ndipo mphete iliyonse ya peptide imakhala ndi zotsalira za amino acid pafupifupi 67 mpaka 75.Kapangidwe ndi kakonzedwe ka ma amino acid m'chigawo cha V kumatsimikizira mtundu wa antigen womanga wa antibody.Chifukwa cha mitundu yosinthika nthawi zonse komanso kutsatizana kwa ma amino acid mdera la V, mitundu yambiri ya ma antibodies okhala ndi ma antigen osiyanasiyana amatha kupangidwa.Magawo a V a unyolo wa L ndi H unyolo amatchedwa VL ndi VH, motsatana.Mu VL ndi VH, kapangidwe ka amino acid ndi kutsatizana kwa zigawo zina zakumaloko kumakhala ndi kusiyanasiyana kwakukulu.Madera amenewa amatchedwa hypervariable regions (HVR).Mapangidwe a amino acid ndi makonzedwe a magawo omwe si a HVR m'chigawo cha V ndi osamala, omwe amatchedwa chigawo cha framework.Pali zigawo zitatu zosinthika mu VL, zomwe nthawi zambiri zimakhala zotsalira za amino acid 24 mpaka 34 ndi 89 mpaka 97 motsatana.Ma HVR atatu a VL ndi VH amatchedwa HVR1, HVR2 ndi HVR3, motsatana.Kafukufuku ndi kusanthula kwa X-ray crystal diffraction yatsimikizira kuti dera losasinthika ndilodi malo omwe antigen antigen amamangirira, motero amatchedwa complementarity-determining region (CDR).HVR1, HVR2 ndi HVR3 ya VL ndi VH imatha kutchedwa CDR1, CDR2 ndi CDR3 motsatana.Nthawi zambiri, CDR3 ili ndi digiri yapamwamba ya hypervariability.Dera la hypervariable ndiyenso malo akuluakulu omwe ma idiotypic determinants of Ig molecules alipo.Nthawi zambiri, unyolo wa H umagwira ntchito yofunika kwambiri pomanga antigen.
Dera lokhazikika (C dera)ili pa 3/4 kapena 4/5 (pafupifupi kuchokera ku amino acid 119 mpaka C terminal) ya H unyolo pafupi ndi C terminus ndi 1/2 (ili ndi zotsalira za 105 za amino acid) pafupi ndi C terminus ya L unyolo.Chigawo chilichonse chogwira ntchito cha unyolo wa H chimakhala ndi zotsalira za 110 za amino acid, ndipo zimakhala ndi mphete ya peptide yopangidwa ndi zotsalira za amino acid 50-60 zolumikizidwa ndi zomangira za disulfide.Mapangidwe a amino acid ndi makonzedwe a dera lino ndizokhazikika mumtundu womwewo wa Ig isotype L ndi mtundu womwewo wa H.Momwemonso, imatha kumangirira mwachindunji ku antigen yofananira, koma mawonekedwe a dera lake C ndi ofanana, ndiko kuti, ali ndi antigenicity yofanana.Kavalo wotsutsana ndi munthu wa IgG wachiwiri (kapena wotsutsa-anti-antibody) akhoza kuphatikizidwa ndi awiri A kuphatikiza ma antibodies (IgG) motsutsana ndi ma exotoxins osiyanasiyana amapezeka.Ichi ndi maziko ofunikira pokonzekera ma antibodies achiwiri ndikugwiritsa ntchito fluorescein, isotopes, michere ndi ma antibodies ena olembedwa.
 
 
Zogwirizana nazo:
Cell Direct RT-qPCR zida