Name: personalized peptide arrays for detection of hla alloantibodies in organ transplantation
Uploaded: 2017-09-06T15:00:00
Description: Watch this Scientific Journal Video about Personalized Peptide Arrays for Detection of HLA Alloantibodies in Organ Transplantation at JoVE.com

Vi takker Drs. Shawn Li og Xing Li fra Western University i Canada for deres venlige assistance med SPOT array-produktion. Vi er taknemmelige for ansatte i kernen Histocompatibility og på den omfattende hårtransplantation Center af Northwestern University prøve tjenesteydelser. Dette arbejde er blevet delvist støttet af hjælpeansatte bestyrelsen af det nordvestlige Memorial Hospital, og et fakultet start fond fra Northwestern University til J.J..

Alle metoder beskrevet her er blevet godkendt ved Northwestern University institutionelle Review Board (IRB protokol #: STU00104680). En overordnet arbejdsproces i protokollen er illustreret i figur 1.
 1. Bioinformatic analyse af Donor og Recipient HLA sekvenser <ol><li>hente sekvenser fra IMGT/HLA database 15.
	<ol><li>Få HLA skrive rapporter organdonor og hans/hendes recipients. 
		Bemærk: Sikre passende procedurer til at beskytte...

<ol>
	<li>Bradshaw, R. A., Dunn, P. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Unambiguous+high+resolution+genotyping+of+human+leukocyte+antigens.">Unambiguous high resolution genotyping of human leukocyte antigens.</a> J Immunol Methods. , (2017).</li><li>Robinson, J., Halliwell, J. A., McWilliam, H., Lopez, R., Marsh, S. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=IPD--the+Immuno+Polymorphism+Database.">IPD--the Immuno Polymorphism Database.</a> Nucleic Acids Res. 41, D1234-D1240 (2013).</li><li>Tait, B. 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A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=HLA+antibody+detection+with+solid+phase+assays:+great+expectations+or+expectations+too+great?.">HLA antibody detection with solid phase assays: great expectations or expectations too great?.</a> Am J Transplant. 14 (9), 1964-1975 (2014).</li><li>Horton, R., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Gene+map+of+the+extended+human+MHC.">Gene map of the extended human MHC.</a> Nat Rev Genet. 5 (12), 889-899 (2004).</li><li>Duquesnoy, R. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=HLAMatchmaker:+a+molecularly+based+algorithm+for+histocompatibility+determination+I.+Description+of+the+algorithm.">HLAMatchmaker: a molecularly based algorithm for histocompatibility determination I. Description of the algorithm.</a> Hum Immunol. 63 (5), 339-352 (2002).</li><li>Duquesnoy, R. J., Marrari, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=HLAMatchmaker-based+definition+of+structural+human+leukocyte+antigen+epitopes+detected+by+alloantibodies.">HLAMatchmaker-based definition of structural human leukocyte antigen epitopes detected by alloantibodies.</a> Curr Opin Organ Transplant. 14 (4), 403-409 (2009).</li><li>Wedel, J., Bruneau, S., Kochupurakkal, N., Boneschansker, L., Briscoe, D. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Chronic+allograft+rejection:+a+fresh+look.">Chronic allograft rejection: a fresh look.</a> Curr Opin Organ Transplant. 20 (1), 13-20 (2015).</li><li>Rostaing, L. P., Malvezzi, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=HLA-Incompatible+Kidney+Transplantation--Worth+the+Risk?.">HLA-Incompatible Kidney Transplantation--Worth the Risk?.</a> N Engl J Med. 374 (10), 982-984 (2016).</li><li>Gebel, H. M., Bray, R. A., Nickerson, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Pre-transplant+assessment+of+donor-reactive,+HLA-specific+antibodies+in+renal+transplantation:+contraindication+vs.+risk.">Pre-transplant assessment of donor-reactive, HLA-specific antibodies in renal transplantation: contraindication vs. risk.</a> Am J Transplant. 3 (12), 1488-1500 (2003).</li><li>Haas, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=An+updated+Banff+schema+for+diagnosis+of+antibody-mediated+rejection+in+renal+allografts.">An updated Banff schema for diagnosis of antibody-mediated rejection in renal allografts.</a> Curr Opin Organ Transplant. 19 (3), 315-322 (2014).</li><li>Cecka, J. M., Reed, E. F., Zachary, A. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=HLA+high-resolution+typing+for+sensitized+patients:+a+solution+in+search+of+a+problem?.">HLA high-resolution typing for sensitized patients: a solution in search of a problem?.</a> Am J Transplant. 15 (4), 855-856 (2015).</li><li>Tambur, A. R., Claas, F. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=HLA+epitopes+as+viewed+by+antibodies:+what+is+it+all+about?.">HLA epitopes as viewed by antibodies: what is it all about?.</a> Am J Transplant. 15 (5), 1148-1154 (2015).</li><li>Robinson, J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+IPD+and+IMGT/HLA+database:+allele+variant+databases.">The IPD and IMGT/HLA database: allele variant databases.</a> Nucleic Acids Res. 43, D423-D431 (2015).</li><li>Gabriel, C., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=HLA+typing+by+next-generation+sequencing+-+getting+closer+to+reality.">HLA typing by next-generation sequencing - getting closer to reality.</a> Tissue Antigens. 83 (2), 65-75 (2014).</li><li>Liu, P., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+Novel+Method+for+Anti-HLA+Antibody+Detection+Using+Personalized+Peptide+Arrays.">A Novel Method for Anti-HLA Antibody Detection Using Personalized Peptide Arrays.</a> Transplant Direct. 2 (11), (2016).</li><li>Frank, R., Overwin, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=SPOT+synthesis.+Epitope+analysis+with+arrays+of+synthetic+peptides+prepared+on+cellulose+membranes.">SPOT synthesis. Epitope analysis with arrays of synthetic peptides prepared on cellulose membranes.</a> Methods Mol Biol. 66, 149-169 (1996).</li><li>Frank, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+SPOT-synthesis+technique.+Synthetic+peptide+arrays+on+membrane+supports--principles+and+applications.">The SPOT-synthesis technique. Synthetic peptide arrays on membrane supports--principles and applications.</a> J Immunol Methods. 267 (1), 13-26 (2002).</li><li>Amartely, H., Iosub-Amir, A., Friedler, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Identifying+protein-protein+interaction+sites+using+peptide+arrays.">Identifying protein-protein interaction sites using peptide arrays.</a> J Vis Exp. (93), (2014).</li><li>Kudithipudi, S., Kusevic, D., Weirich, S., Jeltsch, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Specificity+analysis+of+protein+lysine+methyltransferases+using+SPOT+peptide+arrays.">Specificity analysis of protein lysine methyltransferases using SPOT peptide arrays.</a> J Vis Exp. (93), e52203 (2014).</li><li>Hilpert, K., Winkler, D. F., Hancock, R. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Peptide+arrays+on+cellulose+support:+SPOT+synthesis,+a+time+and+cost+efficient+method+for+synthesis+of+large+numbers+of+peptides+in+a+parallel+and+addressable+fashion.">Peptide arrays on cellulose support: SPOT synthesis, a time and cost efficient method for synthesis of large numbers of peptides in a parallel and addressable fashion.</a> Nat Protoc. 2 (6), 1333-1349 (2007).</li><li>McBride, R., Head, S. R., Ordoukhanian, P., Law, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Low-Cost+Peptide+Microarrays+for+Mapping+Continuous+Antibody+Epitopes.">Low-Cost Peptide Microarrays for Mapping Continuous Antibody Epitopes.</a> Methods Mol Biol. 1352, 67-83 (2016).</li><li>Li, S. S., Wu, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Using+peptide+array+to+identify+binding+motifs+and+interaction+networks+for+modular+domains.">Using peptide array to identify binding motifs and interaction networks for modular domains.</a> Methods Mol Biol. 570, 67-76 (2009).</li><li>Kaczmarek, I., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Donor-specific+HLA+alloantibodies:+long-term+impact+on+cardiac+allograft+vasculopathy+and+mortality+after+heart+transplant.">Donor-specific HLA alloantibodies: long-term impact on cardiac allograft vasculopathy and mortality after heart transplant.</a> Exp Clin Transplant. 6 (3), 229-235 (2008).</li><li>Claas, F. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Clinical+relevance+of+circulating+donor-specific+HLA+antibodies.">Clinical relevance of circulating donor-specific HLA antibodies.</a> Curr Opin Organ Transplant. 15 (4), 462-466 (2010).</li><li>Brown, N. K., Kheradmand, T., Wang, J., Marino, S. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Identification+and+characterization+of+novel+HLA+alleles:+Utility+of+next-generation+sequencing+methods.">Identification and characterization of novel HLA alleles: Utility of next-generation sequencing methods.</a> Hum Immunol. 77 (4), 313-316 (2016).</li><li>Cino, E. A., Choy, W. Y., Karttunen, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Conformational+Biases+of+Linear+Motifs.">Conformational Biases of Linear Motifs.</a> Journal of Physical Chemistry B. 117 (50), 15943-15957 (2013).</li><li>Duquesnoy, R. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Human+leukocyte+antigen+epitope+antigenicity+and+immunogenicity.">Human leukocyte antigen epitope antigenicity and immunogenicity.</a> Curr Opin Organ Transplant. 19 (4), 428-435 (2014).</li><li>Lavinder, J. 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Design af matrixen SPOT beskrevet her er for eksperimentel undersøgelse af alloantibody specificitet i transplantation mod en organdonor HLA antigener. I modsætning til den eksisterende SAB assay bredt brugt i klinikken, har antigen array metode en stor fordel i sin fleksible design, der kan rumme de sande HLA sekvenser af den enkelte donor. Den nye platform udnytter potentialerne af den hastige teknologiske DNA-sekventering teknologi, der vil snart være i stand til at producere præcise HLA allel sekvens aflæsninger...

Orgel substitutionsbehandling, der rutinemæssigt gennemføres i hele verden har reddet millioner af menneskeliv. Solid organtransplantation forekommer i ca. 100 patienter pr. million indbyggere i USA hvert år, mens et større antal stadig er på ventelister til at modtage donor organer på grund af alvorlige forsyningsmangel (ifølge oplysninger fra orglet Udtagning og Transplantation netværket - OPTN/Uno: optn.transplant.hrsa.gov). Organtransplantation er stærkt reguleret for at reducere orgel affald og redde liv, men de videnskabelige værktøjer bruges til at informere disse forordninger er begrænset i effektivitet. For eksempel er det videnskabelige samfund anerkender fuldt ud de meget polymorfe stater af molekyler, HLA og nøjagtige genetiske tests af DNA ved hjælp af høj opløsning maskinskrivning og sekvens-baseret skrive (SBT) er blevet udviklet i de seneste år1, 2., alloantibody testmetoder har endnu ikke stand til at producere de mange forskellige individuelle HLA sekvenser som antigen sonder. Den standard test i dag bruger en uforanderlig panel af ~ 100 allel antigener, der er sammensat af almindelige varianter af HLA, A, B, C, DQ, DP og DR sekvenser i befolkningsgrupper3,4,5,6. Ofte, den faktiske donor HLA sekvenser er ikke inkluderet i panelet test, tvinger transplantation læger og kirurger til at udlede donor-specifikke reaktivitet baseret på delte ligheder mellem donor's faktiske sekvenser og tilsvarende "standarder" i den test sæt7,8. Det er derfor, nogle gange udfordrende at foretage et pålideligt skøn over afvisning risikobaseret antistof test resultater9,10,11,12. Nye personligt tilpasselig tests for alloantibodies er derfor tvingende nødvendige13,14.
HLA-gener indkode store histocompatibility complex (MHC) receptorer, der har en nøglefunktion i immunrespons6. HLA gener kendt for at være de mest polymorfe gener af det menneskelige genom6. På grund af de hurtige fremskridt i DNA sekventering strategier for HLA-gener, nye allel varianter (eller blot benævnt alleler) bliver opdaget på en eksplosiv hastighed15,16. I marts 2017, 16,755 validerede alleler havde deponeret til IMGT/HLA-databasen (http://www.ebi.ac.uk/ipd/index.html), af hvilke 12,351 blev af klasse og 4,404 var af klasse II grupper. I skarp kontrast, er kun lidt over 100 forskellige alleler repræsenteret i standard enkelt-antigen perler (SAB) assay, der rutinemæssigt anvendes til at registrere alloantibodies i organtransplantation. Metoden SAB er bygget på en Luminex platform ved hjælp af flowcytometri. Eftersom analysen udnytter et ufravigeligt sæt af antigener, bortset fra mindre batch til batch variabilitet i produktion, kan antiserum test standardiseret håndfast, på tværs af individer og laboratorier5. Denne test er imidlertid ikke i stand til at fange alle alloantibodies udviklet specifikt mod donor alleler, især når donor sekvenser er fraværende fra SAB sæt. Selv om brugerdefinerede produktion af donorernes antigener baseret på sande sekvenser er ønskelig, fortsat der tekniske udfordringer i strømline den nødvendige produktion og testprocedurer.
Vi beskrev for nylig en alternativ metode i en gennemførlighedsundersøgelse af renal transplantation fag17. Metoden anvendes peptid antigener i en array format til sondering før og efter transplantationen sera af enkelte fag. Hver matrix var brugerdefineret bygget ved hjælp af SPOT syntese metode18,19,20,21,22,23 , producerer peptid antigener, hver 15 aminosyrer i længden, udelukkende baseret på de respektive organdonor HLA alleler af A, B, C, DQA1, DQB1 og DRB1. SPOT syntese drives på en cellulose membran ved hjælp af standard Fmoc-kemi22 og kan producere hundredvis af brugerdefinerede peptider parallelt med en fuldt automatiseret robotsystem19,21. Matrixen membran kan modstå flere runder af stripning og reprobing cykler. I vores retrospektiv undersøgelse17registreret vi ændringer i antigen mønstre med lagrede transplantation antisera indsamlet i en tidsserie (dvs. før og efter transplantation). Heri beskriver vi de teknisk protokol for den arbejdsproces, herunder array design, fremstilling, antiserum sondering og resultat analyse. Metoden er beregnet til påvisning af alloantibodies mod specifikke lineære epitoper på transplantation donorernes HLA molekyler.

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			<td height="40" style="height:40px;width:167px;">Ethanol</td>
			<td style="width:141px;">Sigma-Aldrich</td>
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			<td style="width:141px;">Sigma-Aldrich</td>
			<td style="width:115px;">P7170</td>
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			<td height="40" style="height:40px;width:167px;">Non-fat milk</td>
			<td style="width:141px;">Bio Rad Laboratories</td>
			<td style="width:115px;">1706404</td>
			<td style="width:155px;"></td>
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			<td height="40" style="height:40px;width:167px;">TBST</td>
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			<td style="width:115px;">10711454001</td>
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			<td height="40" style="height:40px;width:167px;">Goat anti-human IgG&#8211;HRP&#160;</td>
			<td style="width:141px;">ThermoFisher Scientific</td>
			<td style="width:115px;">A18811</td>
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			<td height="40" style="height:40px;width:167px;">Clarity Western ECL Substrate</td>
			<td style="width:141px;">Bio Rad Laboratories</td>
			<td style="width:115px;">1705061</td>
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			<td height="40" style="height:40px;width:167px;">Restore Western Blot Stripping Buffer</td>
			<td style="width:141px;">Thermo Scientifics</td>
			<td style="width:115px;">21059</td>
			<td style="width:155px;"></td>
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			<td height="40" style="height:40px;width:167px;">ChemiDoc gel imaging system</td>
			<td style="width:141px;">Bio Rad Laboratories</td>
			<td style="width:115px;">1708265&#160;</td>
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personalized peptide arrays for detection of hla alloantibodies in organ transplantation

I organtransplantation stole funktion og lang levetid af graften kritisk på succesen af kontrollerende immunologiske afvisning reaktivitet mod menneskets leukocyt antigen (HLA). Histocompatibility retningslinjer er baseret på laboratorieundersøgelser af anti-HLA immunitet, der præsenterer enten som præ-eksisterende eller de novo genereret HLA antistoffer, der udgør en større transplantation barriere. Aktuelle prøver er bygget på en single-antigen perler (SAB) platform ved hjælp af et fast sæt af ~ 100 forudvalgt rekombinant HLA antigener for at afprøve transplant sera. Men, hos mennesker der findes en langt større vifte af HLA typer, med ingen to personer identiske tvillinger, der kan dele den samme kombination af HLA sekvenser. Mens avancerede teknologier til HLA skrivning og direkte sekvensering kan netop fange eventuelle uoverensstemmelser i DNA sekvens mellem en donor og modtagerens HLA, SAB assay, på grund af dens begrænsede udvalg i sekvens repræsentation, ikke er i stand til at præcist registrere alloantibodies specielt mod donor HLA uoverensstemmelser. Vi forsøgte at udvikle en supplerende metode bruger en anden teknologi til at opdage og karakterisere anti-donor HLA antistoffer på personlig basis. Værktøjet screening er en brugerdefineret peptid vifte af donor HLA-afledte sekvenser for sondering efter transplantation sera af orgel modtageren til at vurdere risikoen for antistof-medieret afvisning. På en enkelt array for et giver / modtager par, er op til 600 unikke peptider foretaget på grundlag af donors HLA protein sekvenser, hver peptid transporterer mindst én uoverensstemmende rester i en 15-amino syre sekvens. I vores pilotforsøg at sammenligne antigen mønstre for før og efter hårtransplantation sera på disse arrays, var vi i stand til at opdage anti-HLA signaler med den beslutning, som også tilladt os at lokalisere den immun epitoper, der er involveret. Disse personlige antigen arrays tillade høj opløsning påvisning af donor-specifikke HLA epitoper i organtransplantation.

I den oprindelige undersøgelse ved hjælp af matrixen screening metode17, indskrevet vi ialt 5 nyrer transplant emner. Vi opnåede HLA at skrive resultatet af vores kohorte og deres respektive donorer. Deres sygehistorie og allel antistof titers fra SAB tests var også tilgængelige for os. I vores pilot-undersøgelse af disse 5 patienter, vi udtænkt to forskellige metoder: en standard array, består af et fast panel af peptider og personlig arrays, der var skik ...

Watch this Scientific Journal Video about Personalized Peptide Arrays for Detection of HLA Alloantibodies in Organ Transplantation at JoVE.com

Personalized Peptide Arrays for Detection of HLA Alloantibodies in Organ Transplantation

Uoverensstemmelser i human leukocyt antigen (HLA) sekvenser mellem organdonor og modtagende par er den største årsag til antistof-medieret afvisning i organtransplantation. Her præsenterer vi brugen af brugerdefinerede antigen arrays, der er baseret på individuelle donorer HLA sekvenser at sonde anti-donor HLA alloantibodies i orgel modtagere.

Personlig peptid Arrays for påvisning af HLA Alloantibodies i organtransplantation

In organ transplantation, the function and longevity of the graft critically rely on the success of controlling immunological rejection reactivity against ...

Research

JoVE Journal

Biochemistry

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Easy Manipulation of Architectures in Protein-based Hydrogels for Cell Culture Applications

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Electrochemical Detection of Deuterium Kinetic Isotope Effect on Extracellular Electron Transport in Shewanella oneidensis MR-1

Electrochemical Detection of Deuterium Kinetic Isotope Effect on Extracellular Electron Transport in Shewanella oneidensis MR-1

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Isolering af vævs Ekstrudulære vesikler fra leveren

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Et Fluorogenic peptid kavalergang Assay til at screene for proteolytiske aktivitet: ansøgninger om coronavirus spike protein aktivering

Enveloped viruses such as coronaviruses or influenza virus require proteolytic cleavage of their fusion protein to be able to infect the host cell. Often ...

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Påvisning af proteinubiquitinationssteder ved peptidberigelse og massespektrometri

The posttranslational modification of proteins by the small protein ubiquitin is involved in many cellular events. After tryptic digestion of ...

Study on the Metabolism of Six Systemic Insecticides in a Newly Established Cell Suspension Culture Derived from Tea (Camellia Sinensis L.) Leaves

Study on the Metabolism of Six Systemic Insecticides in a Newly Established Cell Suspension Culture Derived from Tea Camellia Sinensis L. Leaves

Undersøgelse af metabolismen af seks systemiske insekticider i en nyetableret celle affjedrings kultur afledt af te (Camellia sinensis L.) Blade

A platform for studying insecticide metabolism using in vitro tissues of tea plant was developed. Leaves from sterile tea plantlets were induced to form ...