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Qaybaha silica ee daloola ayaa lagu diyaariyey habka sol-gel oo leh qaar ka mid ah wax ka beddelka si loo helo qaybo makroporous ah. Qaybahaas waxaa laga soo saaray by wareejinta silsiladda jajabinta isku-darka ah ee isku-darka ah (RAFT) polymerization with N-phenylmaleimide-methylvinylisocyanate (PMI) iyo styrene si ay u diyaariyaan N-phenylmaleimstyrerrow-polyborerrow. Tiirar bir ah oo aan lahayn (100 × 1.8 mm id) ayaa lagu rakibay xirmo dhoobo ah.Qiimayn PMP tiirka kala soocida peptide isku dar ah oo ka kooban shan peptides (Gly-Tyr, Gly-Leu-Tyr, Gly-Gly-Tyr-Arg, Tyr-Ile-Gly-Ser-Argography ee bini'aadamka trypsin chromat, le) Serum albumin (HAS) . Marka la eego shuruudaha elution-ka ugu fiican, tirada saxanka aragtida ee isku dhafka peptide waa mid aad u sarreeya 280,000 taarikada / m². Marka la barbardhigo waxqabadka kala-soocidda tiirka la sameeyay ee ganacsiga Ascentis Express RP-Amide, waxaa la arkay in waxqabadka kala-soocidda ee tiirka PMP uu ka sarreeyay tiirka xallinta ganacsiga marka la eego effici.
Sanadihii la soo dhaafay, warshadaha biopharmaceutical waxay noqdeen suuq caalami ah oo ballaarinaya iyadoo koror la taaban karo uu ku yimid saamiga suuqa. Iyada oo kobaca qarxa ee warshadaha biopharmaceutical1,2,3, falanqaynta peptides iyo borotiinada ayaa aad loo rabaa borotiinada ku jira dareeraha jidhka, unugyada iyo unugyada waa hawl aad u adag sababtoo ah tirada badan ee noocyada suurtagalka ah ee la ogaan karo ee hal muunad.In kasta oo spectrometry mass ay tahay qalab wax ku ool ah oo loogu talagalay peptide iyo borotiinka isku xigxiga, haddii shaybaarada noocan oo kale ah lagu duro spectrometer mass in hal baas, kala soocida ma noqon doonto mid ku habboon. Dhibaatadani waa la dhimi karaa iyada oo la adeegsanayo falanqaynta dareeraha ee MS ka hor hirgelinta chromography ee falanqeeyayaasha gelaya spectrometer mass ee waqti la siiyay4,5,6. Intaa waxaa dheer, inta lagu jiro kala-soocidda wajiga dareeraha, falanqeeyayaashu waxay diiradda saari karaan gobollada cidhiidhiga ah, taas oo xoogga saaraysa falanqaynta iyo hagaajinta dareenka MS.
Dib-u-celinta dareeraha chromatography (RP-LC) ayaa si ballaaran loogu isticmaalaa nadiifinta iyo kala-soocidda isku-dhafka peptide-ka iyadoo la adeegsanayo octadecyl-modified silica (ODS) sida marxaladda taagan Wajiyada joogtada ah ee loogu talagalay ayaa looga baahan yahay in lagu falanqeeyo peptides iyo borotiinnada leh qaybaha polar iyo non-polar si ay ula falgalaan oo ay u sii hayaan analytes16. Isku-dhafka-mode chromatography, kaas oo bixiya isdhexgalka multimodal, wuxuu noqon karaa beddelka RP-LC ee kala-soocidda peptides, borotiinnada, iyo isku-dhafka kale ee adag. peptide iyo kala-soocidda borotiinka17,18,19,20,21.Habka isku dhafan ee taagan (WAX / RPLC, HILIC / RPLC, polar intercalation / RPLC) waxay ku habboon yihiin kala-soocidda peptide iyo borotiinka sababtoo ah joogitaanka labada kooxood ee polar iyo non-polar22,23,24,25,28,26,26 kooxaha dabaylaha covalently bond waxay muujinayaan awood wanaagsan oo kala soocida iyo xulashada gaarka ah ee falanqaynta dabaysha iyo kuwa aan dabaylaha ahayn, maadaama kala-soociddu ay ku xidhan tahay isdhexgalka u dhexeeya falanqaynta iyo marxaladda taagan. Isdhexgalka Multimodal 29, 30, 31, 32. Dhawaan, Zhang et al. 30 waxay diyaarisay dodecyl-la joojiyay marxaladda taagan ee polyamine waxayna si guul leh u kala soocday hydrocarbons, antidepressants, flavonoids, nucleosides, estrogens, iyo dhowr analytes. tiirarka) ayaa ganacsi ahaan lagu heli karaa iyada oo hoos timaada magaca ganacsiga Ascentis Express RP-Amide columns, laakiin tiirarkan waxaa loo isticmaalaa falanqaynta amin 33 kaliya.
Daraasadda hadda jirta, wejiga joogtada ah ee cirifka ah (N-phenylmaleimide-embedded polystyrene) ayaa la diyaariyey oo lagu qiimeeyay kala-soocidda peptides iyo dheef-shiidka trypsin ee HSA. Wajiga taagan ayaa la diyaariyey iyadoo la adeegsanayo istaraatiijiyada soo socota (PEG), TMOS, biyaha acetic acid ayaa la hagaajiyay si loo diyaariyo qaybo silica leh size pore weyn.Second, ligand cusub, phenylmaleimide-methyl vinyl isocyanate, ayaa la farsameeyey oo loo isticmaalo in derivatize qaybaha silica si ay u diyaariyaan a polar ah gundhig taagan stage.The natiijada marxalad idman taagan yahay columned 1 × ah.0 Nidaamka baakadaha (Gly-Tyr, Gly-Leu-Tyr, Gly-Gly-Tyr-Arg, Tyr-Ile-Gly-Ser-Arg, Leucine Enkephalin) iyo Trypsin dheefshiidka ee serum albumin aadanaha (HAS). Tiirka RP-Amide.Labada peptides iyo borotiinada ayaa loo arkay inay si fiican u xalliyeen oo ay ugu hufan yihiin tiirka PMP, kaas oo ka waxtar badnaa tiirka Ascentis Express RP-Amide.
PEG (Polyethylene Glycol), Urea, Acetic Acid, Trimethoxy Orthosilicate (TMOS), Trimethyl Chlorosilane (TMCS), Trypsin, Serum Albumin (HSA), Ammonium Chloride, Urea, Hexane Methyldisilazane (HMDS), Methacryloyl Chloride (MC), Styroxy-H4, Peroxide (BPO), HPLC Grade Acetonitrile (ACN), Methanol, 2-Propanol, iyo Acetone laga soo iibsaday Sigma-Aldrich (St. Louis, MO, USA).
Isku dar ah HOL (8 g), polyetylen glycol (8 g), iyo 8 mL oo ah 0.01 N acetic acid ayaa la walaaqay 10 daqiiqo, ka dibna 24 mL oo TMOS ah ayaa lagu daray xaaladaha baraf-qabow. Isku darka falcelinta ayaa lagu kululeeyay 40 ° C 6 saacadood ka dibna 120 ° C biyo nadiif ah ayaa lagu shubay 8 saacadood oo biyo ah. waxaa la qalajiyey 70 ° C saacadaha 12. Cufnaanta jilicsan ee la qalajiyey waxay ahayd dhul siman oo foorno ah oo lagu dhajiyay 550 ° C saacadaha 12. Saddex qaybood ayaa la diyaariyey waxaana lagu gartaa in lagu baaro dib-u-soo-nooleynta xajmiga qaybta, xajmiga daloolka iyo aagga dusha sare.
Beddelka dusha sare ee qaybaha silica leh ligand phenylmaleimide-methylvinylisocyanate (PCMP) oo ay ku xigto radial polymerization oo leh styrene, koox koox-kooxeed ah ayaa la diyaariyey. Marxaladda taagan ee isku-darka iyo silsiladaha polystyrene. Habka diyaarinta ayaa lagu qeexay hoos.
N-phenylmaleimide (200 mg) iyo methyl vinyl isocyanate (100 mg) ayaa lagu milmay toluene qalalan, iyo 0.1 mL ee 2,2'-azoisobutyronitrile (AIBN) ayaa lagu daray weelka falcelinta si loogu diyaariyo phenylmaleimide-methyl vinyl isocyanate copolymer. la sifeeyay oo lagu qalajiyey foornada 40 ° C ilaa 3 saacadood.
Qaybaha silica qalalan (2 g) ayaa lagu kala firdhiyey toluene qalalan (100 mL), walaaqay oo lagu sonicated 500 mL wareega hoose ee galaaska 10 min.PMCP (10 mg) ayaa lagu milmay toluene waxaana lagu daray dropwise si fallaadho falcelin ah iyada oo loo marayo funnel daadinta 60 ° C saacadaha 3. Kadibna, qaybaha silica ee PMCP-ku-xidhan (100 g) ayaa lagu milmay toluene (200 ml) iyo 4-hydroxy-TEMPO (2 mL) ayaa lagu daray iyadoo ay joogaan 100 µL of dibutyltin dilaure sida kicinta. Isku-darka ayaa lagu walaaqay saacadaha 80 ° C.
Styrene (1 mL), benzoyl peroxide BPO (0.5 mL), iyo TEMPO-PMCP-ku-xiran qaybaha silica (1.5 g) ayaa lagu kala firdhiyey toluene waxaana lagu nadiifiyaa nitrogen.
Shaybaarada waxaa lagu dhajiyay 393 K saacada 1 si loo helo cadaadiska hadhaaga ah ee ka yar 10-3 Torr. Qadarka N2 ee lagu dhajiyay cadaadiska qaraabada ah ee P / P0 = 0.99 ayaa loo isticmaalay si loo go'aamiyo wadarta guud ee mugga daloolka. The morphology of bare and ligand-bonded silica particles (High microscopy Japan) (silica bare iyo ligand-bonded silica particles) ayaa lagu dhejiyay tiir aluminium ah iyadoo la adeegsanayo cajalad kaarboon ah oo dhejis ah.Gold ayaa lagu dhejiyay shaybaarada iyadoo la adeegsanayo koodhka Q150T, iyo lakabka 5 nm Au ayaa lagu shubay shaybaarada. Tani waxay hagaajinaysaa waxtarka habka iyadoo la adeegsanayo koronto yar oo bixiya hadhuudh fiican, spultham qabow, USA Electron. EA1112 Elemental analyzer ayaa loo isticmaalay falanqaynta aasaasiga ah.A Malvern (Worcestershire, UK) Mastersizer 2000 falanqeeyaha cabbirka qayb ayaa loo isticmaalay si loo helo qaybinta qaybinta qaybinta. Kursiga Mastersizer. Falanqaynta Thermogravimetric waxaa lagu sameeyay heerkul ah 5 °C daqiiqadii in ka badan heerkul ah 30 ilaa 800 °C.
Dhalada ku dahaaran tiirarka cidhiidhiga ah ee birta dhuuban ee cabbirka (100 × 1.8 mm id) ayaa la soo buuxiyay iyadoo la adeegsanayo habka xirxirida qulqulka, iyadoo la adeegsanayo isla nidaamka loo adeegsaday Ref. 31.Tiir bir ah oo aan lahayn (glass-lined, 100 × 1.8 mm id) oo leh meel bannaan oo ku habboon oo ka kooban 1 µm frit ayaa lagu xidhay baakad slurry ah (Alltech Deerfield, IL, USA) . Diyaari slurry weji taagan adigoo joojinaya 150 mg ee safka safka ah ee mL iyo u dir kaydinta methan2 mL 1 met. Waxaa loo adeegsaday sida dareeraha dareeraha ah iyo sidoo kale dareeraha kicinta. Buuxi tiirka si isdaba joog ah adigoo isticmaalaya cadaadiska 100 MP ee 10 daqiiqo, 80 MP 15 daqiiqo, iyo 60 MP ee 30 daqiiqo. Inta lagu jiro xirmada, gariir farsamo ayaa lagu dabaqay laba ruxruxayaal GC ah (Alltech, Deerfield, IL, IL, IL) si loo sii daayo xirmooyinka lebiska. Cadaadiska si tartiib tartiib ah si looga hortago waxyeello kasta oo ka mid ah tiirka dhexdiisa. Ka saar tiirarka cutubka baakadaha dareeraha ah oo ku xidh mid kale oo ku habboon gudaha gudaha iyo nidaamka LC si aad u hubiso waxqabadkiisa.
Bamka LC (10AD Shimadzu, Japan), injector (Valco (USA) C14 W.05) oo leh 50nL duritaanka loop, xuubka degasser (Shimadzu DGU-14A), daaqada koofiyadda UV-VIS ayaa la dhisay Qalabka µLC Gaarka ah (UV-2075) iyo isku xidhka muraayadaha-suubyada yaryar ee dhuuban ee dheeraadka ah. baakooyinka ka dib, capillaries (50 μm id 365 iyo yaraynta capillaries midowga (50 μm) ayaa lagu rakibay 1/16 "outlet ee ururka yaraynta. Xog ururinta iyo habaynta chromatographic waxaa lagu sameeyey iyadoo la isticmaalayo software Multichro 2000. Kormeerka at 254 nm Analytes waxaa lagu tijaabiyey Chromat xogta. (Northampton, MA).
Albumin laga bilaabo serum bini'aadamka, budada lyophilized, ≥ 96% (agarose gel electrophoresis) 3 mg oo lagu qaso trypsin (1.5 mg), 4.0 M urea (1 ml), iyo 0.2 M ammonium bicarbonate (1 mL) 0.1% TFA. Sifee xalka oo ku kaydi wax ka hooseeya 4 °C.
Kala soocida isku dhafka peptide iyo dheefshiidka HSA trypsin ayaa si gaar ah loogu qiimeeyay tiirarka PMP. Hubi kala soocida isku darka peptide iyo trypsin ee HSA ee column PMP oo barbar dhig natiijada safka Ascentis Express RP-Amide. Tirada saxanka aragtida waxaa loo xisaabiyaa sida soo socota:
Sawirada SEM ee qaybaha silica qaawan iyo qaybaha silica ee isku xidhan ayaa lagu muujiyay FIG. 2. Sawirada SEM ee qaybaha silica qaawan (A, B) waxay muujinayaan in, si ka duwan daraasadaheenii hore, qaybahani waa wareegsan yihiin kuwaas oo qaybuhu ay dheer yihiin ama ay leeyihiin calaamad aan caadi ahayn. qaybaha silica.
Sawirada mikroskoob ee elektarooniga ah ee walxaha silica qaawan (A, B) iyo qaybaha silica-ku-xidhan ee isku xidhan (C, D).
Qaybinta cabbirka qaybinta qaybaha silica qaawan iyo qaybaha silica ee ligand-ku-xidhan ayaa lagu muujiyay Jaantuska 3 (A). of PMP waa 3.36 μm, marka la barbar dhigo our daraasadii hore ee ad (0.5) qiimaha 3.05 μm (polystyrene-xidheen silica particles)34. Tani waxay lahayd qayb qaybinta qayb cidhiidhi ah marka la barbar dhigo our daraasadii hore ay sabab u tahay saamiga kala duwan ee PEG, urea, TMOS, iyo PEG Acid ee qaybta ka weyn ee falcelinta ka weyn ee qaybta Acetic. Marxaladda walxaha silica-ku-xidhan ee polystyrene-ku-xidhan ee aynu hore u barannay. Tani waxay ka dhigan tahay in shaqeynta dusha sare ee qaybaha silica ee leh styrene ay kaliya dhigeen lakabka polystyrene (0.97 µm) dusha silica, halka wajiga PMP uu dhumucdiisu ahayd 1.38 µm.
Qaybinta cabbirka walxaha (A) iyo qaybinta cabbirka daloolka (B) ee qaybinta silica qaawan iyo qaybaha silica-ku-xidhan ee ligand.
Cabbirka daloolada, mugga daloolka iyo dusha sare ee qaybaha silica ee daraasadda hadda jirta ayaa lagu bixiyaa Shaxda 1 (B) . Noocyada PSD ee qaybaha silica ee qaawan iyo ligand-bonded silica particles ayaa lagu muujiyay sawirka 3 (B) Natiijadu waxay la mid tahay daraasaddeenii hore waxay muujinaysaa in xajmiga daloolka uu hoos u dhaco 69 ka dib markii wax laga beddelo kiimikaad, sida ku cad Shaxda 1 (B), iyo isbedelka qalooca ayaa lagu muujiyay sawirka 3 (B) Daraasaddeenii hore (124 m2 / g) .Sida ku cad shaxda 1 (B), aagga dusha sare (m2 / g) ee qaybaha silica ayaa sidoo kale hoos u dhacay 116 m2 / g ilaa 105 m2 / g ka dib isbeddelka kiimikada.
Natiijooyinka falanqaynta aasaasiga ah ee marxaladda taagan ayaa lagu muujiyay shaxda 2. Xamuulka kaarboonka ee marxaladda taagan ee hadda taagan waa 6.35%, taas oo ka hooseysa culeyska kaarboonka ee daraasaddeena hore ( polystyrene bonded silica particles, 7.93% 35 iyo 10.21%, siday u kala horreeyaan) styrene, qaar ka mid ah ligands polar sida phenylmaleimide-methylvinylisocyanate (PCMP) iyo 4-hydroxy-TEMPO ayaa la isticmaalay.Miisaanka nitrogen ee boqolkiiba marxaladda taagan hadda waa 2.21%, marka la barbardhigo 0.1735 iyo 0.85% by miisaanka nitrogen ee daraasadihii hore, siday u kala horreeyaan. phenylmaleimide.Similarly, loading carbon ee alaabta (4) iyo (5) ahaayeen 2.7% iyo 2.9%, siday u kala horreeyaan, halka loading carbon ee alaabta kama dambaysta ah (6) ahaa 6.35%, sida ku cad Shaxda 2. The miisaanka luminta waxaa lagu hubiyaa marxaladda taagan ee PMP, iyo qalooca TGA ayaa lagu muujiyay sawirka 8. Xamuulka kaarboonka (6.35%) sababtoo ah xargaha waxa ku jira oo kaliya C laakiin sidoo kale N, O, iyo H.
The phenylmaleimide-methylvinylisocyanate ligand ayaa loo doortay wax ka beddelka dusha sare ee qaybaha silica sababtoo ah waxay leedahay kooxaha phenylmaleimide polar iyo kooxaha vinylosyanate.Vinyl isocyanate group.Vinyl isocyanate groups more falcelin la styrene by noolaha polymerization xagjirka ah.Sababta labaad waa in la geliyo koox leh isdhexgalka dhexdhexaadka ah oo aan lahayn isdhexgal dhexdhexaad ah oo u dhexeeya analy station. wajiga, tan iyo moiety phenylmaleimide ma laha lacag dalwaddii at pH.The polarity ee marxaladda taagan waxaa lagu xakameyn karaa qadarka ugu fiican ee styrene iyo waqtiga falcelinta ee polymerization.The ugu dambeeya ee falcelinta (polymerization-free xagjirka ah) waa mid muhiim ah oo bedeli kara polarity ee marxaladda taagan. Falanqaynta Elemental ayaa la sameeyay si loo hubiyo in heerka kor u kacaya ee kaarboonka ayaa la arkay. iyo waqtiga falcelintu waxay kordhisay loading carbon ee marxaladda taagan iyo lidkeeda.SPs loo diyaariyey la uruursanay kala duwan ee styrene leeyihiin loadings carbon kala duwan. Mar labaad, load kuwaas wejiyada taagan ee tiirar birta ahama iyo hubi waxqabadkooda chromatographic (doorashada, xallinta, qiimaha N, iwm) .. Iyada oo ku saleysan tijaabooyinkan, qaabaynta la hagaajiyay ayaa la doortay si loo hubiyo in PMP ay dib u soo celinayso marxaladda saxda ah ee la xakameeyey.
Shan isku dhaf ah oo peptide ah (Gly-Tyr, Gly-Leu-Tyr, Gly-Gly-Tyr-Arg, Tyr-Ile-Gly-Ser-Arg, leucine enkephalin) ayaa sidoo kale lagu qiimeeyay iyadoo la adeegsanayo tiirka PMP iyadoo la adeegsanayo marxalad wareeg ah; 60/40 (v / v) acetonitrile / biyaha (0.1% TFA) ee heerka socodka 80 μL / min. Marka la eego shuruudaha ugu fiicnaanta, lambarka saxanka aragtida (N) ee tiirarka (100 × 1.8 mm id) waa 20,000 ± 100 (200,000 P) ee qiimaha 3. tiirarka iyo chromatogram-yada ayaa lagu muujiyay Jaantuska 5A. Falanqaynta degdega ah ee tiirka PMP ee heerka socodka sare (700 μL / min), shan peptides ayaa la soo saaray hal daqiiqo gudaheed, N qiyamka ayaa ahaa mid aad u wanaagsan, 13,500 ± 330 halkii tiir (100 × 1.8 mm id), Waxay u dhigantaa 135, cabir ahaan 00 saxan. tiirarka (100 × 1.8 mm id) waxaa lagu soo buuxiyey saddex qaybood oo kala duwan oo PMP ah oo taagan si loo hubiyo reproducibility. Falanqaynta falanqaynta ee tiir kasta ayaa la duubay iyada oo la adeegsanayo xaaladaha ugu habboon ee tayada iyo tirada taarikada aragtida N iyo wakhtiga haynta si loo kala saaro isku dhafka tijaabada ah ee tiir kasta. Xogta taranka ee tiirarka PMP ayaa lagu muujiyay shaxda dib-u-celinta ee si fiican u hooseeya. % qiimayaasha RSD, sida ku cad Shaxda 3.
Kala soocida isku dhafka peptide ee tiirka PMP (B) iyo Ascentis Express RP-Amide column (A); wajiga mobaylka 60/40 ACN/H2O (TFA 0.1%), cabbirada tiirka PMP (100 × 1.8 mm id); Analytical Nidaamka elution ee xeryahooda: 1 (Gly-Tyr), 2 (Gly-Leu-Tyr), 3 (Gly-Gly-Tyr-Arg), 4 (Tyr-Ile-Gly-Ser-Arg) iyo 5 (leucine) acid enkephalin)).
Tiirka PMP (100 × 1.8 mm id) ayaa lagu qiimeeyay kala soocida dheefshiidka tryptic ee serum albumin ee waxqabadka sare ee chromatography dareeraha. TFA.Sida lagu muujiyay chromatogram-ka (Jaantuska 6), dheef-shiid kiimikaadka HSA ayaa loo kala qaybiyay 17 ugu sarreeya oo u dhiganta 17 peptides. Waxtarka kala-soocidda ee mid kasta oo ugu sarreeya ee dheef-shiid kiimikaadka HSA ayaa la xisaabiyay waxaana qiimaha lagu bixiyaa shaxda 5.
Dheefshiidka tijaabinta ee HSA (100 × 1.8 mm id) ayaa lagu kala soocay tiirka PMP; heerka socodka (100 µL/min), wareega wareega 60/40 acetonitrile/biyo leh 0.1% TFA.
halka L waa dhererka tiirarka, η waa viscosity ee wajiga wareega, ΔP waa cadaadiska dhabarka ee tiirka, iyo u waa xawaaraha toosan ee marxaladda wareega wareega wareega wareega ee PMP wuxuu ahaa 2.5 × 10-14 m2, heerka qulqulka wuxuu ahaa 25 μL / min, iyo 60/40 v / v ACN / biyaha waxaa loo adeegsaday 1 × 0. id) waxay la mid ahayd tii daraasaddeenii hore ee Ref.34. Dareemida tiirka ay ka buuxaan qaybo dusha sare ka daloola waa: 1.7 × 10-15 oo ah 1.3 μm qaybo, 3.1 × 10-15 oo ah 1.7 μm, 5.2 × 10-15 . Qaybaha 5 μm Qaybaha 43. Sidaa darteed, qulqulka wajiga PMP wuxuu la mid yahay 5 μm qaybaha qolofka xudunta u ah.
halka Wx uu yahay miisaanka tiirka lagu kaxayn karo chloroform, Wy waa miisaanka tiirka ka buuxa methanol, iyo ρ waa cufnaanta dareeraha. Cufnaanta methanol (ρ = 0.7866) iyo chloroform (ρ = 1.484) . wadarta guud ee porosity ee SILICA PARTICLES 3 × 1 × 0. Tiirarka C18-Urea 31 ee aan horay u barannay waxay ahaayeen 0.63 iyo 0.55, siday u kala horreeyaan Qaybaha silica sababtoo ah marxaladaha joogtada ah ee C18-nooca C18 waxay ku xiran yihiin qaybaha silica sida silsiladaha toosan, halka polystyrene-nooca marxaladaha taagan, lakabka polymer qaro weyn ayaa laga sameeyay hareeraha.
Jaantuska 7A,B wuxuu muujinayaa tiirka PMP (100 × 1.8 mm id) iyo Ascentis Express RP-Amide tiirka (100 × 1.8 mm id) iyadoo la adeegsanayo xaalado isku mid ah (ie, 60/40 ACN/H2O iyo 0.1% TFA). ) ee shirqoolkii van Deemter. Isku darka peptide ee la xushay (Gly-Tyr, Gly-Leu-Tyr, Gly-Gly-Tyr-Arg, Tyr-Ile-Gly-Ser-Arg, Leucine Enkephalin) ayaa lagu diyaariyay 20 µL Tiirka iyo tiirka Ascentis Express RP-Amide waxay ahaayeen 2.6 µm iyo 3.9 µm, siday u kala horreeyaan.Qiimaha HETP waxay muujinayaan in waxtarka kala soocida tiirka PMP (100 × 1.8 mm id) uu aad uga fiican yahay ganacsiga la heli karo Ascentis Express RP-Amide column (100 × 1) Fig. in hoos u dhaca qiimaha N ee qulqulka sii kordhaya uusan ahayn mid muhiim ah marka loo eego daraasaddeennii hore. Waxtarka sare ee kala-soocidda ee tiirka PMP (100 × 1.8 mm id) marka la barbar dhigo tiirka Ascentis Express RP-Amide waxay ku saleysan tahay hagaajinta qaabka qaybta, cabbirka, iyo hababka xirmooyinka cufan ee adag ee loo isticmaalo shaqada hadda34.
(A). ACN/H2O oo leh 0.1% TFA.
A polar-embedded polystyrene wajiga taagan ayaa la diyaariyey oo lagu qiimeeyay kala soocida isku dhafka peptide synthetic iyo trypsin dheefshiidka ee serum albumin aadanaha (HAS) in chromatography dareere waxqabadka sare. size ee qaybaha silica ah, synthesis la xakameeyey ee wejiga taagan, iyo packing.In kakan ee waxtarka kala soocidda sare, column hoose cadaadis dhabarka heerka socodka sare waa faa'iido kale oo this weji. dhirta daawada iyo fangaska laga soosaaray chromatography dareere ah. Mustaqbalka, tiirarka PMP ayaa sidoo kale lagu qiimeyn doonaa kala-soocidda borotiinka iyo unugyada monoclonal.
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