Name: in vitro methods for comparing target binding and cdc induction between therapeutic antibodies: applications in biosimilarity analysis
Uploaded: 2017-05-04T13:00:00
Description: Watch this Scientific Journal Video about In Vitro Methods for Comparing Target Binding and CDC Induction Between Therapeutic Antibodies: Applications in Biosimilarity Analysis at JoVE.com

Forfatterne har ingen bekreftelser.

1. Evaluering av målbinding ved hjelp av flytcytometri <ol><li> Fremstilling av biologiske materialer og reagenser <ol><li> Lag 500 ml RPMI-dyrkningsmedium tilsatt 10% varmeinaktivert føtalt bovint serum (H-IFBS). </li><li> Kultur Daudi Burkitts lymfom (Daudi) celler og Daudi GFP + celler ved bruk av RPMI og 75 cm 2 kulturflasker. Opprettholde kulturer ved 37 ° C i en 5% CO 2 fuktig atmosfære til de når 6 - 9 x 10 5 celler / ml. </li><li> Lag 50 ml farg...

<ol>
	<li>Schimizzi, G. F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Biosimilars+from+a+practicing+rheumatologist+perspective:+An+overview.">Biosimilars from a practicing rheumatologist perspective: An overview.</a> Autoimmun Rev. 15 (9), 911-916 (2016).</li><li>Cuello, H. A., 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=Comparability+of+Antibody-Mediated+Cell+Killing+Activity+Between+a+Proposed+Biosimilar+RTXM83+and+the+Originator+Rituximab.">Comparability of Antibody-Mediated Cell Killing Activity Between a Proposed Biosimilar RTXM83 and the Originator Rituximab.</a> Bio Drugs. 30 (3), 225-231 (2016).</li><li>Iwamoto, N., 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=Validated+LC/MS+Bioanalysis+of+Rituximab+CDR+Peptides+Using+Nano-surface+and+Molecular-Orientation+Limited+(nSMOL)+Proteolysis.">Validated LC/MS Bioanalysis of Rituximab CDR Peptides Using Nano-surface and Molecular-Orientation Limited (nSMOL) Proteolysis.</a> Biol Pharm Bull. 39 (7), 1187-1194 (2016).</li><li>Chapman, 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=Waiving+in+vivo+studies+for+monoclonal+antibody+biosimilar+development:+National+and+global+challenges.">Waiving in vivo studies for monoclonal antibody biosimilar development: National and global challenges.</a> MAbs. 8 (3), 427-435 (2016).</li><li>Zembruski, N. 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=7-Aminoactinomycin+D+for+apoptosis+staining+in+flow+cytometry.">7-Aminoactinomycin D for apoptosis staining in flow cytometry.</a> Anal Biochem. 429 (1), 79-81 (2012).</li><li>Salinas-Jazmin, N., Hisaki-Itaya, E., Velasco-Velazquez, M. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+flow+cytometry-based+assay+for+the+evaluation+of+antibody-dependent+cell-mediated+cytotoxicity+(ADCC)+in+cancer+cells.">A flow cytometry-based assay for the evaluation of antibody-dependent cell-mediated cytotoxicity (ADCC) in cancer cells.</a> Methods Mol Biol. 1165, 241-252 (2014).</li><li>Teeling, J. L., 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+Biological+Activity+of+Human+CD20+Monoclonal+Antibodies+Is+Linked+to+Unique+Epitopes+on+CD20.">The Biological Activity of Human CD20 Monoclonal Antibodies Is Linked to Unique Epitopes on CD20.</a> J Immunol. 177 (1), 362-371 (2006).</li><li>Miranda-Hernandez, M. 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=Assessment+of+physicochemical+properties+of+rituximab+related+to+its+immunomodulatory+activity.">Assessment of physicochemical properties of rituximab related to its immunomodulatory activity.</a> J Immunol Res. 2015, 910763 (2015).</li><li>Visser, 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=Physicochemical+and+functional+comparability+between+the+proposed+biosimilar+rituximab+GP2013+and+originator+rituximab.">Physicochemical and functional comparability between the proposed biosimilar rituximab GP2013 and originator rituximab.</a> BioDrugs. 27 (5), 495-507 (2013).</li><li>Ylera, F., 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=Off-rate+screening+for+selection+of+high-affinity+anti-drug+antibodies.">Off-rate screening for selection of high-affinity anti-drug antibodies.</a> Anal Biochem. 441 (2), 208-213 (2013).</li><li>Broyer, L., Goetsch, L., Broussas, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Evaluation+of+complement-dependent+cytotoxicity+using+ATP+measurement+and+C1q/C4b+binding.">Evaluation of complement-dependent cytotoxicity using ATP measurement and C1q/C4b binding.</a> Methods Mol Biol. 988, 319-329 (2013).</li><li>Herbst, 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=B-cell+depletion+in+vitro+and+in+vivo+with+an+afucosylated+anti-CD19+antibody.">B-cell depletion in vitro and in vivo with an afucosylated anti-CD19 antibody.</a> J Pharm Exp Ther. 335 (1), 213-222 (2010).</li><li>Lazar, G. A., 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=Engineered+antibody+Fc+variants+with+enhanced+effector+function.">Engineered antibody Fc variants with enhanced effector function.</a> Proc Natl Acad Sci U S A. 103 (11), 4005-4010 (2006).</li><li>Winiarska, 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=Statins+impair+antitumor+effects+of+rituximab+by+inducing+conformational+changes+of+CD20.">Statins impair antitumor effects of rituximab by inducing conformational changes of CD20.</a> PLoS medicine. 5 (3), e64 (2008).</li><li>Zhou, X., Hu, W., Qin, X. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+role+of+complement+in+the+mechanism+of+action+of+rituximab+for+B-cell+lymphoma:+implications+for+therapy.">The role of complement in the mechanism of action of rituximab for B-cell lymphoma: implications for therapy.</a> Oncologist. 13 (9), 954-966 (2008).</li><li>Hayashi, 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=Gemcitabine+enhances+rituximab-mediated+complement-dependent+cytotoxicity+to+B+cell+lymphoma+by+CD20+upregulation.">Gemcitabine enhances rituximab-mediated complement-dependent cytotoxicity to B cell lymphoma by CD20 upregulation.</a> Cancer Sci. 107 (5), 682-689 (2016).</li><li>Mossner, 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=Increasing+the+efficacy+of+CD20+antibody+therapy+through+the+engineering+of+a+new+type+II+anti-CD20+antibody+with+enhanced+direct+and+immune+effector+cell-mediated+B-cell+cytotoxicity.">Increasing the efficacy of CD20 antibody therapy through the engineering of a new type II anti-CD20 antibody with enhanced direct and immune effector cell-mediated B-cell cytotoxicity.</a> Blood. 115 (22), 4393-4402 (2010).</li><li>Lapalombella, 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=A+novel+Raji-Burkitt's+lymphoma+model+for+preclinical+and+mechanistic+evaluation+of+CD52-targeted+immunotherapeutic+agents.">A novel Raji-Burkitt's lymphoma model for preclinical and mechanistic evaluation of CD52-targeted immunotherapeutic agents.</a> Clin Cancer Res. 14 (2), 569-578 (2008).</li><li>Mitoma, H., 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=Mechanisms+for+cytotoxic+effects+of+anti-tumor+necrosis+factor+agents+on+transmembrane+tumor+necrosis+factor+alpha-expressing+cells:+comparison+among+infliximab,+etanercept,+and+adalimumab.">Mechanisms for cytotoxic effects of anti-tumor necrosis factor agents on transmembrane tumor necrosis factor alpha-expressing cells: comparison among infliximab, etanercept, and adalimumab.</a> Arthritis Rheum. 58 (5), 1248-1257 (2008).</li><li>Kaymakcalan, Z., 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=Comparisons+of+affinities,+avidities,+and+complement+activation+of+adalimumab,+infliximab,+and+etanercept+in+binding+to+soluble+and+membrane+tumor+necrosis+factor.">Comparisons of affinities, avidities, and complement activation of adalimumab, infliximab, and etanercept in binding to soluble and membrane tumor necrosis factor.</a> Clin Immunol. 131 (2), 308-316 (2009).</li><li>Zent, C. S., 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=Direct+and+complement+dependent+cytotoxicity+in+CLL+cells+from+patients+with+high-risk+early-intermediate+stage+chronic+lymphocytic+leukemia+(CLL)+treated+with+alemtuzumab+and+rituximab.">Direct and complement dependent cytotoxicity in CLL cells from patients with high-risk early-intermediate stage chronic lymphocytic leukemia (CLL) treated with alemtuzumab and rituximab.</a> Leuk Res. 32 (12), 1849-1856 (2008).</li><li>Goswami, M. 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=Regulation+of+complement-dependent+cytotoxicity+by+TGF-beta-induced+epithelial-mesenchymal+transition.">Regulation of complement-dependent cytotoxicity by TGF-beta-induced epithelial-mesenchymal transition.</a> Oncogene. 35 (15), 1888-1898 (2016).</li><li>Wang, A., 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=Induction+of+anti-EGFR+immune+response+with+mimotopes+identified+from+a+phage+display+peptide+library+by+panitumumab.">Induction of anti-EGFR immune response with mimotopes identified from a phage display peptide library by panitumumab.</a> Oncotarget. , (2016).</li><li>Ueda, N., 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+cytotoxic+effects+of+certolizumab+pegol+and+golimumab+mediated+by+transmembrane+tumor+necrosis+factor+alpha.">The cytotoxic effects of certolizumab pegol and golimumab mediated by transmembrane tumor necrosis factor alpha.</a> Inflamm Bowel Dis. 19 (6), 1224-1231 (2013).</li><li>Nesbitt, A., 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=Mechanism+of+action+of+certolizumab+pegol+(CDP870):+in+vitro+comparison+with+other+anti-tumor+necrosis+factor+alpha+agents.">Mechanism of action of certolizumab pegol (CDP870): in vitro comparison with other anti-tumor necrosis factor alpha agents.</a> Inflamm Bowel Dis. 13 (11), 1323-1332 (2007).</li><li>Teeling, J. L., 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=Characterization+of+new+human+CD20+monoclonal+antibodies+with+potent+cytolytic+activity+against+non-Hodgkin+lymphomas.">Characterization of new human CD20 monoclonal antibodies with potent cytolytic activity against non-Hodgkin lymphomas.</a> Blood. 104 (6), 1793-1800 (2004).</li></ol>

Patentutløpet av et terapeutisk mAb er å fremme utviklingen av biosimilarer. Dermed er det behov for enkle metoder som kan identifisere forskjeller i klinisk relevante aktiviteter av disse produktene. CD20 + dyrkede celler ble benyttet for evaluering av to sentrale funksjonelle karakteristika ved rituximab: målbinding og CDC-induksjon. Den tidligere aktiviteten krever anerkjennelse av CD20 av Fab-regionen av mAb, mens sistnevnte hovedsakelig avhenger av samspillet mellom Fc-regionen og dets komplement <sup...

Terapeutiske antistoffer rives rekombinante monoklonale antistoffer (mAbs) som er utviklet for behandling av forskjellige sykdommer, inkludert kreft, autoimmune og kroniske sykdommer, nevrologiske lidelser, og andre 1. Foreløpig har FDA gitt godkjenning til mer enn 40 terapeutiske mAbs, og flere er ventet å komme på markedet i de neste årene. Rituximab er et høy-affinitet kimært monoklonalt IgG1 antistoff godkjent for behandling av CD20 + B-celle-non-Hodgkins lymfom (NHL), CD20 + follikulær NHL, kronisk lymfocytisk leukemi, og reumatoid artritt 2, 3. Erkjennelsen av CD20, som er overuttrykt i B-celler, ved rituximab induserer apoptose; komplement aktivering; og antistoffavhengig cellemediert cytotoksisitet (ADCC) 3. Patentene av dette stoffet utløpt i Europa og i USA i 2013 og 2016, Henholdsvis. Dermed utvikler farmasøytiske selskaper over hele verden rituximabbiosimilarer. Som i ethvert annet legemiddel til konsum, krever biosimilarer godkjenning fra reguleringsorganer. Internasjonale retningslinjer indikerer at for mAbs bør biosimilaritet påvises ved å sammenligne de fysisk-kjemiske egenskapene, farmakokinetikken, effekten og sikkerheten til de nye og referanseproduktene 4 . Følgelig må metodene som brukes i slike sammenligninger vurdere de strukturelle og funksjonelle egenskapene til mAbs, spesielt de med klinisk relevans. Til det formål viser in vitro- analyser flere fordeler i forhold til in vivo- eksperimenter (gjennomgått i Chapman et al. ) 5 : i) in vitro- studier er mer sensitive for forskjeller mellom de foreslåtte biosimilar og referanseproduktet; Ii) in vivo studier må utføres i relevante arter, som for mange mAbs erikke-humane primater; og iii) siden virkningsmekanisme, den prekliniske toksikologi, og de kliniske virkningene av referanseproduktet er vel kjent in vivo-studier med biosimilars kan ikke gi ytterligere nyttig informasjon. Følgelig EUs Veiledning for biosimilars gjør kandidatene til å gå inn kliniske studier basert på robust in vitro-data alene seks. Her presenterer vi to raske, økonomiske og enkle analyser som evaluerer den biologiske aktiviteten til rituximab ved hjelp av CD20 + dyrkede celler. Disse analysene kan inngå som en del av sammenlignbarhet øvelsen for rituximab biosimilar kandidater.

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>RPMI-1640 medium</td>
			<td>ATCC</td>
			<td>30-2001</td>
			<td>Modify the culture depending on the cell line</td>
		</tr>
		<tr>
			<td>Trypan Blue solution</td>
			<td>Sigma</td>
			<td>T8154</td>
			<td>0.4%, liquid, sterile-filtered, suitable for cell culture</td>
		</tr>
		<tr>
			<td>Daudi Burkitt&#39;s Lymphoma Cells</td>
			<td>ATCC</td>
			<td>CCL-213</td>
			<td>You can modify the cell line depending on the antibody of interest</td>
		</tr>
		<tr>
			<td>Fetal bovine serum(FBS)</td>
			<td>GIBCO</td>
			<td>16000-044</td>
			<td>You can modify the source of serum depending of requirements of the cell line</td>
		</tr>
		<tr>
			<td>Normal Human Serum Complement</td>
			<td>Quidel</td>
			<td>A113</td>
			<td>It is therefore appropriate for use in biocompatibility experiments including drug development, biomaterials testing and other applications</td>
		</tr>
		<tr>
			<td>7AA-D</td>
			<td>BDPharmigen</td>
			<td>559925</td>
			<td>You can use broad range of color options, compatible with most instrument configurations for to analyze viability.</td>
		</tr>
		<tr>
			<td>PECy5 Mouse Anti-human IgG</td>
			<td>BDPharmigen</td>
			<td>551497</td>
			<td>Change fluorochrome depending on the filter and laser of your flow cytometer.</td>
		</tr>
		<tr>
			<td>Human IgG Isotype Control</td>
			<td>ThermoFisher Scientific</td>
			<td>07-7102</td>
			<td>Change depending to mAb</td>
		</tr>
		<tr>
			<td>BDCytofix</td>
			<td>BDPharmigen</td>
			<td>554655</td>
			<td>Flow Cytometry Fixation Buffer (1-4% formaldehyde or paraformaldehyde )</td>
		</tr>
		<tr>
			<td>PBS pH 7.4 10X (Phosphate buffer saline)</td>
			<td>GIBCO</td>
			<td>70011-044</td>
			<td>Phosphatebuffer without Ca2+/Mg2+ [137 mM NaCl, 2.7 mM KCl, 8 mM Na2HPO4, 1.46 mM KH2PO4] and endotoxin free.</td>
		</tr>
		<tr>
			<td>Cell culture plates 96 well, V-bottom</td>
			<td>Corning</td>
			<td>29442-068</td>
			<td>12 x 75 mm round bottom test tubes or 96-well V- or U- bottom microtiter plates</td>
		</tr>
		<tr>
			<td>MabThera (Rituximab)</td>
			<td>Roche</td>
			<td>&#8212;</td>
			<td>Reference product</td>
		</tr>
		<tr>
			<td>Rituximab</td>
			<td>Indian</td>
			<td>&#8212;</td>
			<td>Biosimilar product</td>
		</tr>
		<tr>
			<td>15- or 50-mL conical centrifuge tubes</td>
			<td>Corning</td>
			<td>430290 or 430052</td>
			<td>&#8212;</td>
		</tr>
		<tr>
			<td>Pipette Tips</td>
			<td>Eppendorf</td>
			<td>&#8212;</td>
			<td>Multiple volume configurations are necessary</td>
		</tr>
		<tr>
			<td>Pipettes</td>
			<td>Eppendorf</td>
			<td>&#8212;</td>
			<td>Adjustable-volume pipettes are necessary</td>
		</tr>
		<tr>
			<td>Centrifuge 5430/ 5430R model</td>
			<td>Eppendorf</td>
			<td>&#8212;</td>
			<td>Refrigerated variable-speed centrifuge (4 to 25 &#176; C) with speeds ranging from 10 to 30,130 &#215; g</td>
		</tr>
		<tr>
			<td>Flow cytometer</td>
			<td>BD Dickinson</td>
			<td>&#8212;</td>
			<td>BD FACSAria III or other flow cytometer</td>
		</tr>
		<tr>
			<td>Olympus optical and light microscope</td>
			<td>Olympus</td>
			<td>&#8212;</td>
			<td>To quantify and evaluate cell growth</td>
		</tr>
		<tr>
			<td>Incubator</td>
			<td>SANYO</td>
			<td>&#8212;</td>
			<td>Incubatorfor temperature andCO2 control to culture cells</td>
		</tr>
		<tr>
			<td>Biological Safety Cabinet</td>
			<td>CHC BIOLUS</td>
			<td>&#8212;</td>
			<td>Biological safety cabinet that is used to protect the researcher, product and environment.</td>
		</tr>
	</tbody>
</table>

in vitro methods for comparing target binding and cdc induction between therapeutic antibodies: applications in biosimilarity analysis

Terapeutiske monoklonale antistoffer (mAbs) er relevant ved behandling av forskjellige sykdommer, inkludert kreft. Utviklingen av biosimilar mAbs av farmasøytiske selskaper er en markedsmulighet, men det er også en strategi for å øke narkotika tilgjengelighet og redusere behandlingsrelaterte kostnader. Protokollene som beskrevet her beskriver evalueringen av target-binding og CDC-induksjon med rituximab i Daudi-celler. Disse to funksjonene krever forskjellige strukturelle regioner av antistoff og er relevante for den kliniske virkning indusert av rituximab. Protokollene side-til-side-sammenligning av et referanse rituximab og et markedsført rituximab biosimilar. De evaluerte produktene viste forskjeller både i target-binding og CDC induksjon, noe som antyder at det er underliggende fysikalsk-kjemiske forskjeller, og fremhever behovet for å analysere virkningen av disse forskjeller i klinisk sammenheng. De metoder beskrevet her utgjør enkel og billig in vitro &lt;/ Em&gt; modeller for evaluering av aktiviteten til rituximabbiosimilarer. Dermed kan de være nyttige under biosimilar utvikling, så vel som for kvalitetskontroll i biosimilar produksjon. Videre kan de fremstilte metoder ekstrapoleres til andre terapeutiske mAbs.

Ved hjelp av de protokoller som er beskrevet ovenfor, mål-binding og CDC induksjon av referanse rituximab ble sammenlignet i parallell med de til en biosimilar rituximab produsert og kommersielt tilgjengelig i Asia. I Daudi-celler, både mAbs bundet CD20 på en konsentrasjonsavhengig måte (figur 1D). Ikke-lineære regresjoner av bindingsdataene viste en r 2 på 0,978 og 0,848 for referanse og bios...

Watch this Scientific Journal Video about In Vitro Methods for Comparing Target Binding and CDC Induction Between Therapeutic Antibodies: Applications in Biosimilarity Analysis at JoVE.com

In Vitro Methods for Comparing Target Binding and CDC Induction Between Therapeutic Antibodies: Applications in Biosimilarity Analysis

Denne protokollen beskriver in vitro-sammenligning av to viktige funksjonelle egenskaper rituximab: målbindingsdomene og komplementavhengig cytotoksisitet (CDC) induksjon. Metodene som ble benyttet for en side-til-side-sammenligning mellom referanse rituximab rituximab og en biosimilar. Disse analyser kan anvendes under biosimilar utvikling eller som en kvalitetskontroll i sin produksjon.

In vitro- metoder for sammenligning av målbinding og CDC-induksjon mellom terapeutiske antistoffer: applikasjoner i biosimilaritetsanalyse

Therapeutic monoclonal antibodies (mAbs) are relevant to the treatment of different pathologies, including cancers. The development of biosimilar mAbs by ...

Research

JoVE Journal

Immunology and Infection

In Vitro Methods for Comparing Target Binding and CDC Induction Between Therapeutic Antibodies: Applications in Biosimilarity Analysis

Human T Lymphocyte Isolation, Culture and Analysis of Migration In Vitro

Human T Lymphocyte Isolation, Culture and Analysis of Migration In Vitro

The migration of T lymphocytes involves the adhesive interaction of cell surface integrins with ligands expressed on other cells or with extracellular ...

Introducing Shear Stress in the Study of Bacterial Adhesion

During bacterial infections a sequence of interactions occur between the pathogen and its host. Bacterial adhesion to the host cell surface is often the ...

Establishing a Liquid-covered Culture of Polarized Human Airway Epithelial Calu-3 Cells to Study Host Cell Response to Respiratory Pathogens In vitro

Establishing a Liquid-covered Culture of Polarized Human Airway Epithelial Calu-3 Cells to Study Host Cell Response to Respiratory Pathogens In vitro

Etablering av en flytende dekket Culture of Polarized Menneskelig Airway Epiteliale Calu-3 Cells å studere Host Cell Response til luftveispatogener<em&gt; In vitro</em

The  apical and basolateral surfaces of airway epithelial cells demonstrate  directional responses to pathogen exposure in  vivo. Thus, ideal in vitro ...

Total Protein Extraction and 2-D Gel Electrophoresis Methods for Burkholderia Species

Total Protein Extraction and 2-D Gel Electrophoresis Methods for Burkholderia Species

Totalt Protein Extraction og 2-D-gel-elektroforese Metoder<em&gt; Burkholderia</em&gt; Arter

The investigation of  the intracellular protein levels of bacterial species is of importance to  understanding the pathogenic mechanisms of diseases ...

A Novel In vitro Model for Studying the Interactions Between Human Whole Blood and Endothelium

A Novel In vitro Model for Studying the Interactions Between Human Whole Blood and Endothelium

A Novel<em&gt; In vitro</em&gt; Modell for å studere samspillet mellom humant fullblod og endotelet

The majority of all known diseases are accompanied by disorders of the cardiovascular system. Studies into the complexity of the interacting pathways ...

Neutrophil Isolation and Analysis to Determine their Role in Lymphoma Cell Sensitivity to Therapeutic Agents

Nøytrofile Isolering og analyse for å fastslå deres rolle i lymfom Cell Følsomhet for Terapeutiske midler

Neutrophils are the most abundant (40% to 75%) type of white blood cells and among the first inflammatory cells to migrate towards the site of ...

An In Vitro Caseum Binding Assay that Predicts Drug Penetration in Tuberculosis Lesions

An In Vitro Caseum Binding Assay that Predicts Drug Penetration in Tuberculosis Lesions

en<em&gt; In Vitro</em&gt; Caseum bindingsassay som forutsier medikamentpenetrering i Tuberkulose Lesjoner

The eradication of tuberculosis disease requires drug regimens that can penetrate the multiple layers of complex pulmonary lesions. Drug distribution in ...

Induction of Paralysis and Visual System Injury in Mice by T Cells Specific for Neuromyelitis Optica Autoantigen Aquaporin-4

Induksjon av lammelse og visuelle systemet skade i mus ved T-celler spesifikke for Neuromyelitis Optica Autoantigen Aquaporin-4

While it is recognized that aquaporin-4 (AQP4)-specific T cells and antibodies participate in the pathogenesis of neuromyelitis optica (NMO), a human ...

Alternative In Vitro Methods for the Determination of Viral Capsid Structural Integrity

Alternative In Vitro Methods for the Determination of Viral Capsid Structural Integrity

Alternative In Vitro -metoder for fastsettelse av Viral kapsid konstruksjonssikkerhet

Human norovirus exacts considerable public health and economic losses worldwide. Emerging in vitro cultivation advances are not yet applicable for routine ...

Human Colonoid Monolayers to Study Interactions Between Pathogens, Commensals, and Host Intestinal Epithelium

Menneskelig Colonoid Monolayers å studere interaksjonen mellom patogener, Commensals og vert Intestinal epitel

Human 3-dimensional (3D) enteroid or colonoid cultures derived from crypt base stem cells are currently the most advanced ex vivo model of the intestinal ...