Araştırma sözleşmesi Miguel Servet (ISCIII CD14 / 00032) (GN-G.). Bilimsel Araştırmalar &quot;Translational Neuroscience Programı Enstitüsü&quot; Hollanda Teşkilatı (022005019) (CH) den Kardeşliği. Prinses Beatrix Fonds (Proje WAR08-12) ve Ortaklık Française contre les Miyopatiler için hibeler (PM-M.); yanı sıra Hollanda Bilimsel Araştırma Örgütü bir Veni Bursu (916.10.148) Hollanda Beyin Vakfı burs ile (FS2008 (1) -28) ve ML Prinses Beatrix Fonds (Proje WAR08-12) ( ). Biz flow sitometri göre sıralama B-hücresi ona yardım için Jozien Jaspers teşekkür ederiz.

<ol>
	<li>Vrolix, K., 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=Clonal+heterogeneity+of+thymic+B+cells+from+early-onset+myasthenia+gravis+patients+with+antibodies+against+the+acetylcholine+receptor.">Clonal heterogeneity of thymic B cells from early-onset myasthenia gravis patients with antibodies against the acetylcholine receptor.</a> J Autoimmun. 52, 101-112 (2014).</li><li>Yamashita, M., Katakura, Y., Shirahata, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Recent+advances+in+the+generation+of+human+monoclonal+antibody.">Recent advances in the generation of human monoclonal antibody.</a> Cytotechnology. 55 (2-3), 55-60 (2007).</li><li>Pereira, K. M., Dellavance, A., Andrade, L. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+challenge+of+identification+of+autoantibodies+specific+to+systemic+autoimmune+rheumatic+diseases+in+high+throughput+operation:+Proposal+of+reliable+and+feasible+strategies.">The challenge of identification of autoantibodies specific to systemic autoimmune rheumatic diseases in high throughput operation: Proposal of reliable and feasible strategies.</a> Clin Chim Acta. 437, 403-410 (2014).</li><li>Losen, M., 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=Treatment+of+myasthenia+gravis+by+preventing+acetylcholine+receptor+modulation.">Treatment of myasthenia gravis by preventing acetylcholine receptor modulation.</a> Ann N Y Acad Sci. 1132, 174-179 (2008).</li><li>Smith, K., 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=Rapid+generation+of+fully+human+monoclonal+antibodies+specific+to+a+vaccinating+antigen.">Rapid generation of fully human monoclonal antibodies specific to a vaccinating antigen.</a> Nat Protoc. 4 (3), 372-384 (2009).</li><li>Fraussen, 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=A+novel+method+for+making+human+monoclonal+antibodies.">A novel method for making human monoclonal antibodies.</a> J Autoimmun. 35 (2), 130-134 (2010).</li><li>Traggiai, E., 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=An+efficient+method+to+make+human+monoclonal+antibodies+from+memory+B+cells:+potent+neutralization+of+SARS+coronavirus.">An efficient method to make human monoclonal antibodies from memory B cells: potent neutralization of SARS coronavirus.</a> Nat Med. 10 (8), 871-875 (2004).</li><li>Fraussen, 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=Autoantigen+induced+clonal+expansion+in+immortalized+B+cells+from+the+peripheral+blood+of+multiple+sclerosis+patients.">Autoantigen induced clonal expansion in immortalized B cells from the peripheral blood of multiple sclerosis patients.</a> J Neuroimmunol. 261 (1-2), 98-107 (2013).</li><li>Yamashita, M., 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=Different+individual+immune+responses+elicited+by+in+vitro+immunization.">Different individual immune responses elicited by in vitro immunization.</a> Cytotechnology. 40 (1-3), 161-165 (2002).</li><li>Hui-Yuen, J., Koganti, S., Bhaduri-McIntosh, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Human+B+cell+immortalization+for+monoclonal+antibody+production.">Human B cell immortalization for monoclonal antibody production.</a> Methods Mol Biol. 1131, 183-189 (2014).</li><li>Tiller, T., 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=Efficient+generation+of+monoclonal+antibodies+from+single+human+B+cells+by+single+cell+RT-PCR+and+expression+vector+cloning.">Efficient generation of monoclonal antibodies from single human B cells by single cell RT-PCR and expression vector cloning.</a> J Immunol Methods. 329 (1-2), 112-124 (2008).</li><li>Lanzavecchia, A., Corti, D., Sallusto, F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Human+monoclonal+antibodies+by+immortalization+of+memory+B+cells.">Human monoclonal antibodies by immortalization of memory B cells.</a> Curr Opin Biotechnol. 18 (6), 523-528 (2007).</li><li>Traggiai, E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Immortalization+of+human+B+cells:+analysis+of+B+cell+repertoire+and+production+of+human+monoclonal+antibodies.">Immortalization of human B cells: analysis of B cell repertoire and production of human monoclonal antibodies.</a> Methods Mol Biol. 901, 161-170 (2012).</li><li>Strober, W., 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=Monitoring+cell+growth.+Curr+Protoc+Immunol+/+edited+by.">Monitoring cell growth. Curr Protoc Immunol / edited by.</a> John E. Coligan ... [et al.]. Appendix 3, Appendix 3A (2001).</li><li>Ibrahim, S. F., van den Engh, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Flow+cytometry+and+cell+sorting.">Flow cytometry and cell sorting.</a> Adv Biochem Eng Biotechnol. 106, 19-39 (2007).</li><li>Leith, J. T., Padfield, G., Faulkner, L. E., Quinn, P., Michelson, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effects+of+feeder+cells+on+the+X-ray+sensitivity+of+human+colon+cancer+cells.">Effects of feeder cells on the X-ray sensitivity of human colon cancer cells.</a> Radiother Oncol. 21 (1), 53-59 (1991).</li><li>Hui-Yuen, J., McAllister, S., Koganti, S., Hill, E., Bhaduri-McIntosh, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Establishment+of+Epstein-Barr+virus+growth-transformed+lymphoblastoid+cell+lines.">Establishment of Epstein-Barr virus growth-transformed lymphoblastoid cell lines.</a> J Vis Exp. (57), 3321 (2011).</li><li>Smith, L. M., 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=Fluorescence+detection+in+automated+DNA+sequence+analysis.">Fluorescence detection in automated DNA sequence analysis.</a> Nature. 321 (6071), 674-679 (1986).</li></ol>

Yazarlar hiçbir rakip mali çıkarlarını olduğunu beyan ederim.

Bu yazıda, insanlara özgü PBMCs&#39;den elde IgG antikorlarının üretilmesi için tüm adımları ayrıntılı bir şekilde sunulmaktadır. Bu protokol daha önce yayınlanmış tekniklere göre bazı avantajlara sahiptir. Avantajlarından biri üretilen antikor B hücresi klonu orijinal çiftine karşılık gelen ağır ve hafif zincirleri tutmasıdır. IgG antikorlarının tanımlanması insan vericinin her türlü yapılabilir, ve aşı 5 nedeniyle bağışıklık tepkisi alevlenme için bir ihtiyaç ...

Bu maddenin amacı, detaylı bir şekilde oluşturabilir ve insan periferik kan mononükleer hücreleri (PBMC), elde edilen insan IgG monoklonal antikorları karakterize etmek için bir yöntem tanımlamaktır. insan antikorları incelemek için faiz araştırma birçok farklı alanda büyüdü. Özellikle, birçok araştırma grupları oto-antikorların 1-3 kaynaklanan patoloji ilgilendi. Biz klonlanmış ve patojenik oto-antikorlar 1 nitelendirmiştir. oto-antikorların çalışma rakip antikorlar 4 kullanan, örneğin, kendi hedefleri tespit etmek ve tedavi stratejilerinin geliştirilmesine yardımcı olabilir. Ayrıca, insan antikorlarının çalışma aynı zamanda, aşı 5 sonrası bağışıklık tepkisini değerlendirmek için maruz kalan ve belirli bir patojen 6 ya da çalışma için dirençli haline geldi kişilerin antikor profilini karakterize etmek için, örneğin, araştırmanın diğer alanlarda, ilgi hangi Antikorlar olanDoğal repertuar 7,12. Çeşitli teknikler, rekombinant insan monoklonal antikorlarını üretmek için 8-12 geliştirilmiştir; Bunların çoğu, faj görüntüleme ve B hücre immortalizasyonunu kullanımı. Faj gösteriminin kullanımı, geniş ölçüde, yeni antikorlar 13 keşfedilmesi için uygulanmıştır. Ancak haline insan immünoglobulin ağır ve hafif zincir çiftinin işleminde ayrışmış, yani bu, önemli bir dezavantajı vardır. Insan B hücreleri ya da EBV dönüşümü ile hibridomlar üretimi bu dezavantajı ortadan kaldırır. Biz Toll-like reseptör 9 (TLR-9) 6,12&#39;dir yoluyla poliklonal B hücre stimülasyonu ile birlikte EBV ile timik B hücrelerinin enfeksiyonu kullanın. Bu yazıda ayrıntılı olarak in vitro antikor nesil PBMC izolasyondan tüm adımların eksiksiz bir bakış ile, IgG insan antikorlarının gelişmesi için kullanmak teknolojiyi tanımlamak. BuProtokol insan IgG profilin her tür analizi için kullanılabilir. Laboratuvarımızda, IgG antikorları üreten B hücreleri başarıyla sıralama sonra PBMC&#39;lerin geri kalanından ayrılmış oylandı. Elli kriteri B hücreleri 8 daha sonra, çok oyuklu plakalara plakalanır ve bir B hücrelerinin klonal genişleme için, EBV ve TLR 9 aktivasyonu ile immortalize olabilir. Besleyici hücreler olarak, insan embriyonik akciğer dokusundan fibroblastlar, ölümsüzleştirilmiş B hücreleri görüntülenmesini kolaylaştırır hücre çizgisi WI38 kullanılmıştır. Bu B hücreleri, immünoglobulin ağır ve hafif zincirlerinin sekansları, PCR ile elde edilebilir, ve antikor genleri immünoglobülin G sentezleme vektörlerine klonlanmış ve in vitro koşullarda üretilen. Bu tekniği kullanarak, donör bulundu aynı antikor dizisi ile tek antikorlar ele alınabilir.

<table border="0" cellpadding="0" cellspacing="0">
	<tbody>
		<tr>
			<td>Histopaque-1077&#160;</td>
			<td>Sigma-Aldrich</td>
			<td>10771</td>
			<td>solution containing polysucrose and sodium diatrizoate</td>
		</tr>
		<tr>
			<td>FACSAria II cell sorter&#160;</td>
			<td>BD Biosciences&#160;</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>96 U-bottom micro well plates&#160;</td>
			<td>Costar</td>
			<td>3799</td>
			<td></td>
		</tr>
		<tr>
			<td>Advanced Roswell Park Memorial Institute (RPMI) 1640 medium&#160;</td>
			<td>Gibco, Life Technologies</td>
			<td>12633-020</td>
			<td></td>
		</tr>
		<tr>
			<td>30% v/v EBV-containing supernatant of the B95-8 cell line&#160;&#160;</td>
			<td>ATCC</td>
			<td>CRL-1612</td>
			<td>3.4 x 108 copies/ml</td>
		</tr>
		<tr>
			<td>CpG2006&#160;</td>
			<td>Invivogen</td>
			<td>ODN 7909</td>
			<td></td>
		</tr>
		<tr>
			<td>Wi38 cells&#160;</td>
			<td>Sigma-Aldrich</td>
			<td>90020107</td>
			<td></td>
		</tr>
		<tr>
			<td>Interleukin-2</td>
			<td>Roche&#160;</td>
			<td>10799068001</td>
			<td></td>
		</tr>
		<tr>
			<td>ELISA plates</td>
			<td>Greiner Bio-One, Microlon</td>
			<td>655092</td>
			<td></td>
		</tr>
		<tr>
			<td>AffiniPure F(ab&#39;)2 Fragment Goat Anti-Human IgG, Fc&#947; Fragment Specific (unconjugated)</td>
			<td>Jackson ImmunoResearch</td>
			<td>109-006-008</td>
			<td></td>
		</tr>
		<tr>
			<td>4% non-fat dry milk (Blotting Grade Blocker)&#160;</td>
			<td>Biorad</td>
			<td>170-6404</td>
			<td></td>
		</tr>
		<tr>
			<td>Human IgG&#160;</td>
			<td>Sigma</td>
			<td>I 2511</td>
			<td>HUMAN IgG purified Immunoglobulin, 5.6 mg/ml</td>
		</tr>
		<tr>
			<td>Goat F(ab)2 antihuman IgG Fc&#947; (conjugated with peroxidase (PO))</td>
			<td>Jackson ImmunoResearch</td>
			<td>109-036-008</td>
			<td></td>
		</tr>
		<tr>
			<td>ELISA reader (Perkin Elmer 2030)&#160;</td>
			<td>Perkin Elmer&#160;</td>
			<td>2030-0050</td>
			<td></td>
		</tr>
		<tr>
			<td>Peroxidase-conjugated AffiniPure Rabbit Anti-Human IgM, Fc5&#181;&#160;</td>
			<td>Jackson ImmunoResearch</td>
			<td>309-035-095</td>
			<td></td>
		</tr>
		<tr>
			<td>SuperScript III Cells Direct cDNA Synthesis System&#160;</td>
			<td>Invitrogen&#160;</td>
			<td>18080-200</td>
			<td></td>
		</tr>
		<tr>
			<td>Applied Biosystems (ABI) GeneAm PCR System 2700</td>
			<td>Applied Biosystems</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>High Pure RNA Isolation Kit&#160;</td>
			<td>Roche</td>
			<td>11828665001</td>
			<td></td>
		</tr>
		<tr>
			<td>Reverse transcription system kit&#160;</td>
			<td>Promega</td>
			<td>A3500</td>
			<td></td>
		</tr>
		<tr>
			<td>Recombinant Taq DNA Polymerase</td>
			<td>TAKARA</td>
			<td>R001A</td>
			<td></td>
		</tr>
		<tr>
			<td>Primers (2&#956;l)</td>
			<td>Sigma</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Ultrapure Agarose&#160;</td>
			<td>Invitrogen</td>
			<td>16500-500</td>
			<td></td>
		</tr>
		<tr>
			<td>100 bp ladder</td>
			<td>Invitrogen</td>
			<td>15628-019</td>
			<td></td>
		</tr>
		<tr>
			<td>Quantity One 4.5.2 (Gel Doc 2000)</td>
			<td>Biorad</td>
			<td>170-8100</td>
			<td></td>
		</tr>
		<tr>
			<td>QIAquick PCR purification kit</td>
			<td>QIAGEN</td>
			<td>28106</td>
			<td></td>
		</tr>
		<tr>
			<td>BigDye Terminator v3.1 cycle sequencing kit&#160;</td>
			<td>Applied Biosystems</td>
			<td>4337455</td>
			<td></td>
		</tr>
		<tr>
			<td>0.1 ml reaction plate (MicroAMP Optical 96-well)</td>
			<td>Applied Biosystems</td>
			<td>4346906</td>
			<td></td>
		</tr>
		<tr>
			<td>Genetic analyser ABI300&#160;</td>
			<td>Applied Biosystems</td>
			<td>4346906</td>
			<td></td>
		</tr>
		<tr>
			<td>DH5&#945; competent cells (E. coli)</td>
			<td>Invitrogen</td>
			<td>18263-012</td>
			<td></td>
		</tr>
		<tr>
			<td>pFUSEss-CHIg-hG1 (4493 bp)</td>
			<td>Invivogen</td>
			<td>pfusess-hchg1</td>
			<td></td>
		</tr>
		<tr>
			<td>pFUSEss-CHIg-hG4 (4484 bp)&#160;</td>
			<td>Invivogen</td>
			<td>pfusess-hchg4</td>
			<td></td>
		</tr>
		<tr>
			<td>pFUSE2ss-CLIg-hk (3875 bp)</td>
			<td>Invivogen</td>
			<td>pfuse2ss-hclk</td>
			<td></td>
		</tr>
		<tr>
			<td>pFUSE2ss-CLIg-hl2 (3883 bp)&#160;</td>
			<td>Invivogen</td>
			<td>pfuse2ss-hcll2</td>
			<td></td>
		</tr>
		<tr>
			<td>SOC medium</td>
			<td>Invitrogen</td>
			<td>15544-034</td>
			<td></td>
		</tr>
		<tr>
			<td>LB-based agar medium supplemented with Zeocin (Fast-Media Zeo Agar)</td>
			<td>Invivogen</td>
			<td>fas-zn-s</td>
			<td></td>
		</tr>
		<tr>
			<td>Terrific Broth (TB)-based liquid medium supplemented with Zeocin (Fast-Media Zeo TB)</td>
			<td>Invivogen</td>
			<td>fas-zn-l</td>
			<td></td>
		</tr>
		<tr>
			<td>DNA Miniprep kit&#160;</td>
			<td>Omega Bio Technology</td>
			<td>D6942-02</td>
			<td></td>
		</tr>
		<tr>
			<td>Nanodrop (ND1000 Spectrophotometer)</td>
			<td>Nanodrop</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>LB-based agar medium supplemented with Blasticidin (Fast-Media Blast Agar)</td>
			<td>Invivogen</td>
			<td>fas-bl-s</td>
			<td></td>
		</tr>
		<tr>
			<td>Terrific Broth (TB)-based liquid medium supplemented with Blasticidin (Fast-Media Blast TB)</td>
			<td>Invivogen</td>
			<td>fas-bl-l</td>
			<td></td>
		</tr>
		<tr>
			<td>EcoRI</td>
			<td>New England Biolabs</td>
			<td>R0101S</td>
			<td>20,000 U/ml, in 10x NEBuffer EcoRI</td>
		</tr>
		<tr>
			<td>NheI</td>
			<td>New England Biolabs</td>
			<td>R0131S</td>
			<td>10,000 U/ml, in 10x NEBuffer 2.1</td>
		</tr>
		<tr>
			<td>2-Log DNA ladder</td>
			<td>New England Biolabs</td>
			<td>N3200S</td>
			<td>0.1-10.0 kb, 1,000 &#956;g/ml</td>
		</tr>
		<tr>
			<td>XmaI</td>
			<td>New England Biolabs</td>
			<td>R0180S</td>
			<td>10,000 U/ml, in 10x CutSmart Buffer&#160;</td>
		</tr>
		<tr>
			<td>BsiWI</td>
			<td>New England Biolabs</td>
			<td>R0553S</td>
			<td>10,000 U/ml, in 10x NEBuffer 3.1</td>
		</tr>
		<tr>
			<td>AvrII</td>
			<td>New England Biolabs</td>
			<td>R0174S</td>
			<td>5,000 U/ml, in 10x CutSmart Buffer&#160;</td>
		</tr>
		<tr>
			<td>FastAP Thermosensitive Alkaline Phosphatase</td>
			<td>Thermo Scientific</td>
			<td>EF0651</td>
			<td>1 U/&#181;L, in 10x FastAP Buffer</td>
		</tr>
		<tr>
			<td>DH5&#945; competent cells&#160;</td>
			<td>Invitrogen</td>
			<td>18263-012</td>
			<td></td>
		</tr>
		<tr>
			<td>PE Mouse Anti-Human IgG</td>
			<td>BD Pharmingen</td>
			<td>555787</td>
			<td></td>
		</tr>
		<tr>
			<td>anti-CD22, PerCP-Cy5.5, Clone: HIB22</td>
			<td>Fisher scientific</td>
			<td>BDB563942</td>
			<td></td>
		</tr>
		<tr>
			<td>QIAprep Spin Miniprep Kit&#160;</td>
			<td>QIAGEN</td>
			<td>27106</td>
			<td></td>
		</tr>
		<tr>
			<td>BigDye Terminator v3.1</td>
			<td>Applied Biosystems</td>
			<td>4337455</td>
			<td></td>
		</tr>
	</tbody>
</table>

generation of recombinant human igg monoclonal antibodies from immortalized sorted b cells

Finding new methods for generating human monoclonal antibodies is an active research field that is important for both basic and applied sciences, including the development of immunotherapeutics. However, the techniques to identify and produce such antibodies tend to be arduous and sometimes the heavy and light chain pair of the antibodies are dissociated. Here, we describe a relatively simple, straightforward protocol to produce human recombinant monoclonal antibodies from human peripheral blood mononuclear cells using immortalization with Epstein-Barr Virus (EBV) and Toll-like receptor 9 activation. With an adequate staining, B cells producing antibodies can be isolated for subsequent immortalization and clonal expansion. The antibody transcripts produced by the immortalized B cell clones can be amplified by PCR, sequenced as corresponding heavy and light chain pairs and cloned into immunoglobulin expression vectors. The antibodies obtained with this technique can be powerful tools to study relevant human immune responses, including autoimmunity, and create the basis for new therapeutics.

CD22 ve IgG pozitif hücrelerin boyanması sonrası ayırma yolluk, Şekil 1 &#39;de gösterilmiştir, bu görüntü, çift pozitif hücrelerin alanı -. IgG antikorları üreten B hücrelerinin - Ayrı bir tüp içinde bu hücreler sıralamak için seçilir. Analizde, toplam PBMC&#39;lerin yaklaşık% 1&#39;i bu çifte pozitif nüfusa karşılık gelmektedir. Elde edilen kriteri hücrelerin sayısı Bölüm 1 &#39;de elde edilen hücrelerin sayısına bağlıdır. EBV ölü...

Watch this Scientific Journal Video about Generation of Recombinant Human IgG Monoclonal Antibodies from Immortalized Sorted B Cells at JoVE.com

Generation of Recombinant Human IgG Monoclonal Antibodies from Immortalized Sorted B Cells

Synthesis of human monoclonal antibodies is the first step in studies aimed at unraveling the pathophysiological mechanisms of auto-antibody-mediated immune responses. We have developed a protocol to generate recombinant human immunoglobulin G (IgG) monoclonal antibodies from blood sorted B cells, including B-cell isolation, antibody cloning and in vitro synthesis.

Ölümsüzleştirilmiştir Sıralama B hücrelerinden Rekombinant İnsan IgG Monoklonal Antikorlarının Üretimi

Finding new methods for generating human monoclonal antibodies is an active research field that is important for both basic and applied sciences, ...

generation-recombinant-human-igg-monoclonal-antibodies-from

Research

JoVE Journal

Immunology and Infection

18.2K Görüntüleme.  Maastricht University. Synthesis of human monoclonal antibodies is the first step in studies aimed at unraveling the pathophysiological mechanisms of auto-antibody-mediated immune responses. We have developed a protocol to generate recombinant human immunoglobulin G (IgG) monoclonal antibodies from blood sorted B cells, including B-cell isolation, antibody cloning and in vitro synthesis.