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Hybridoma Sequencing Service - Creative Biolabs
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We are committed to providing highly customized comprehensive solutions with the best quality to advance your projects. </p><!--<a href="aboutus.html" class="bannerbutton"><i class="fa fa-angle-right"></i>EXPOLORE MORE</a>--> <b class="Highlights_tit">Highlights</b> <ul class="HighlightsArea"> <li class="col-md-6 col-lg-6 col-sm-6"> <img src="static/images/Abouts-menu-lights-pic-1.jpg" srcset="static/images/Abouts-menu-lights-pic-1.webp 1x" width="200" height="131" loading="lazy"><a href="aboutus.html" class="bannerbutton"> <i class="fa fa-angle-right"></i>About CBL </a> </li> <li class="col-md-6 col-lg-6 col-sm-6"> <img src="static/images/Abouts-menu-lights-pic-2.jpg" srcset="static/images/Abouts-menu-lights-pic-2.webp 1x" width="200" height="131" loading="lazy"><a href="contactus.html" class="bannerbutton"> <i class="fa fa-angle-right"></i>Contact Us </a> </li> </ul> </li> </ul> </div> </div> </li> </ul> <script> window.addEventListener('load',function(){ var arr = ['/drug-discovery/therapeutics/','/bioconjugation/','/adc/']; $('.mennav ul li a').each(function(){ var $this = $(this); var href = $this.attr("href").toLocaleLowerCase(); $.each(arr, function(index, value) { if (href && href.startsWith(value)) { $this.attr('target', '_blank'); } }); }); }); </script> </div> <div class="top-search search-group"> <div class="col-md-5 col-sm-6"> <label class="radio-inline"><input type="radio" name="search" class="rdo_productsearch" value="product" data-type="product" placeholder="Please input cat#, product name, abbr, or keywords."> Product</label> <label class="radio-inline"><input type="radio" name="search" class="rdo_servicesearch" value="service" data-type="service" placeholder="Please input your keywords."> Service</label> <label class="radio-inline"><input type="radio" name="search" class="rdo_googlesearch" value="google" data-type="google" placeholder="Please input your keywords." checked="checked"> Keyword</label> </div> <div class="col-md-7 col-sm-6 search-div"> <input id="keys" name="keys" placeholder="Please input your keywords." class="search-input"> <div class="selectItems"></div> <span class="search_btn"><i class="fa fa-search"></i></span> </div> </div> </div> <div class="searchShow"> <div class="searchtopbg"></div> <span class="searchclosed"><i class="fa fa-times"></i>Close</span> <div class="addsearch search-group"></div> </div> <div class="banner_inpage services_bg_2"> <div class="auto-container small_content-width about_container"> <ul class="bread-crumb clearfix"> <li><a href="/">Home</a></li> <li><a href="javascrpt:void(0)">Characterization</a></li><li><a href="javascrpt:void(0)">Sequence Analysis</a></li> <li>Hybridoma Sequencing</li> </ul> <p>"Creative Biolabs is committed to providing highly customized comprehensive solutions with the best quality to advance our global clients’ projects."</p> </div> </div> <script> window.addEventListener('load', function () { if (($('.bread-crumb li:last-child').text()) == "Peer-Reviewed Publications") { $(".banner_inpage").addClass("peer-reviewed-publications_bg").removeClass("services_bg_2"); } if ($(window).hasClass('bread-crumb')) { if ($(".bread-crumb").html().indexOf("RESOURCE") > -1) { $(".pronote").hide(); } if ($(".bread-crumb").html().indexOf("Careers") > -1) { $(".pronote").hide(); } } }); </script> <div class="small_content-width services-container"> <div class="col-md-9"> <div class="inpage_section_title"> <h1>Hybridoma Sequencing Service</h1> </div> <div class="page-tab"> <span onclick="window.location.href=window.location.pathname+'#tab1'">Background</span> <span onclick="window.location.href=window.location.pathname+'#tab2'">Description</span> <span onclick="window.location.href=window.location.pathname+'#tab3'">Service</span> <span onclick="window.location.href=window.location.pathname+'#tab4'">Highlight</span> <span onclick="window.location.href=window.location.pathname+'#tab5'">Published Data</span> <span onclick="window.location.href=window.location.pathname+'#tab6'">FAQ</span> <span onclick="window.location.href=window.location.pathname+'#tab7'">Resources</span> </div> <script>window.addEventListener('load',function(){page_tab_margin();page_tab_css();})</script> <p> With years of experience in the field of antibody development, <strong>Creative Biolabs</strong> has established a high-accuracy sequencing platform to provide top quality antibody sequence determination services from hybridoma cell lines. Creative Biolabs’ hybridoma sequencing service is highly versatile and is applicable for both IgG and IgM types of monoclonal antibodies produced by mouse and rat hybridoma cell lines. We are dedicated to providing first-class customized service to contribute to the success of your project. </p> <h2 class="h2_tit page_tab_margin" id="tab1"> The Value of Antibody Sequence </h2> <span class="ServiceShowrightPic"><img src="static/img/3-3-Hybridoma-Sequencing-Service-1.jpg" alt="Hybridoma Sequencing Service" width="300" height="228" loading="lazy" /></span> <p> The protein sequence of an antibody is one of its most important attributes that enables various downstream applications and developments: </p> <ol class="ollist"> <li> Patent application: the uniqueness in the amino acid sequence of an antibody, especially that from the variable region, is the key factor that dictates the novelty of the antibody and plays a crucial role in the patent application to ensure the exclusive rights in downstream antibody developments. </li> <li> Antibody recombinant expression: antibody sequence serves as a blueprint for vector construction and recombinant expression in various systems. </li> <li> Antibody downstream development: the original antibody sequence provides a template for various downstream engineering platforms such as humanization and affinity maturation. </li> <li> Other applications: the antibody sequence is also useful in hybridoma cell line validation, antibody format alteration, and to serve as a precaution in case of the loss of hybridoma cell line due to contamination. </li> </ol> <h2 class="h2_tit page_tab_margin" id="tab2"> Hybridoma Sequencing </h2> <p> Hybridoma sequencing refers to the process of obtaining sequence information regarding the cDNA encoding the variable heavy (VH) and variable light (VL) domains of the antibody produced by your hybridoma cell line. Before sequencing, total mRNA of the hybridoma cells is extracted followed by PCR amplification of antibody variable regions (VH and VL) as well as non-variable flanking constant region sequences. The antibody sequence information is derived from the amplified PCR products by a sequencer. With antibody variable region protein sequence, different antibody formats can be designed by grafting the variable region onto various framework sequence templates. Antibodies generated by this method can be produced <em>via</em> recombinant expression in a mammalian expression system. Alternatively, the full-length sequence can also be derived from hybridoma sequencing to enable antibody production <em>via</em> recombinant expression and to serve as an insurance policy to secure the source of the antibody in case of losing the hybridoma cell line due to contamination or other reasons. </p> <p class="ServiceShowPic"> <img src="static/img/3-3-Hybridoma-Sequencing-Service-2.jpg" alt="Schematic representation of the approach for high-throughput sequencing of the Ig sequence repertoire." width="500" height="373" loading="lazy" />Fig 1. Schematic representation of the approach for high-throughput sequencing of the Ig sequence repertoire. (Georgiou, G., 2014) </p> <h2 class="h2_tit page_tab_margin" id="tab3"> Hybridoma sequencing service at Creative Biolabs </h2> <ul class="ullist disc"> <li> Full-length antibody sequencing service </li> </ul> <p> In this service, the 5'RACE method is applied to amplify, clone and sequence the complete sequences of both heavy and light chains of an antibody, which are subsequently cloned into two cloning vectors for downstream sequence manipulation and expression vector constructions. </p> <ul class="ullist disc"> <li> V-region sequencing service </li> </ul> <p> In this service, only the sequences of the heavy and light chains at the variable region of an antibody are amplified by specially designed degenerated primers and sequenced. The resulted variable region can be grafted into any framework to yield an Ig isoform that is most suitable for the client’s project. </p> <ul class="ullist disc"> <li> Recombinant antibody production </li> </ul> <p> With the antibody sequence, scientists at Creative Biolabs provide services to convert the sequences into various forms of antibodies, including multi-isoform full-length antibodies and fragment antibodies such as Fab or scFv. A detailed project report, as well as vectors containing the antibody constructs, are delivered to the client at the end of the project. </p> <h2 class="h2_tit page_tab_margin" id="tab4"> Service Features </h2> <ul class="ullist disc"> <li> The sequencing service is applicable for hybridoma cell lines from most rodent species and antibody isotypes. </li> <li> Highly versatile sequencing services to reveal both variable region and full-length antibody protein sequences. </li> <li> One-stop service from mRNA extraction to sequence determination as well as antibody format conversion. </li> <li> Short turn-around time: the projects are usually finished within 3 weeks. </li> </ul> <br /> <p> Creative Biolabs also offers <a href="/hyperdoma-a-high-throughput-hybridoma-generation-platform.html">high-throughput hybridoma platform</a> and other hybridoma-related services to facilitate your antibody discovery projects. Please feel free to <a href="/antibody-sequencing-service.html#OnlineInquiry">contact us</a> for more information. </p> <p> Other optional <a href="/Antibody-Analysis-Services.html">antibody analysis services</a>: </p> <ul class="ullist"> <li> <a href="/antibody-characterization.html">Antibody Characterization</a> </li> <li> <a href="/Antibody-Affinity-Measurement.html">Antibody Affinity Measurement</a> </li> <li> <a href="/next-generation-antibody-sequencing.html"><em>De Novo</em> Antibody Sequencing Services</a> </li> <li> <a href="/precision-paratope-mapping-service.htm">Precision Paratope Mapping Service</a> </li> <li> <a href="/Monoclonal-Antibody-Epitope-Binning.html">Monoclonal Antibody Epitope Binning</a> </li> <li> <a href="/SPR-Services.html">SPR Services</a> </li> <li> <a href="/Isothermal-Titration-Calorimetry-ITC-Service.html">Isothermal Titration Calorimetry (ITC) Services</a> </li> <li> <a href="/Differential-scanning-calorimetry-DSC-Service.html">Differential Scanning Calorimetry (DSC) Services</a> </li> <li> <a href="/Dual-Polarization-Interferometry-DPI-Services.html">Dual-Polarization Interferometry (DPI) Services</a> </li> </ul> <h2 class="h2_tit page_tab_margin" id="tab5"> Published Data </h2> <ul class="ullist"> <li> <strong>The Development of Humanized Antibodies for CCHFV by Mouse Hybridoma Approach with Subsequent Sequencing</strong> </li> </ul> <p class="show-center"> <img loading="lazy" src="/static/img/1-2-3-3-hybridoma-sequencing-service-2.jpg" width="350" height="190" alt="" />Fig. 2 Model of the EG5 VH and VL domains. (Stuart D. Dowall, 2023) </p> <p> Humanized antibodies to the Crimean-Congo hemorrhagic virus (CCHFV) are essential for the development and standardization of serological tests. The researchers generated humanized monoclonal antibodies against CCHFV through two schemes and compared the two methods. The first is the traditional mouse hybridoma method, followed by sequencing and humanization of the antibody, and the second is a non-animal alternative using the human combinatorial antibody library (HuCAL). The results of the experiment and comparison showed that humanization after hybridoma in mice could produce antibodies with higher affinity. The sequencing data obtained can be used to produce recombinant antibodies in subsequent production and research processes, thus reducing the use of animals for this application in the future. Finally, the researchers provided information about the development of a humanized standardized control, which could be an important positive control component for serological testing of CCHFV. </p> <div class="reference"> <strong>References</strong> <ol class="ollist decimal"> <li> Georgiou, G., (2014). "The promise and challenge of high-throughput sequencing of the antibody repertoire." <em>Nature biotechnology</em>, 32(2), 158-168. </li> <li> Dowall, Stuart D., et al. "Development of humanised antibodies for Crimean-Congo Haemorrhagic fever virus: Comparison of hybridoma-based versus phage library techniques." <em>Journal of Immunological Methods</em> 512 (2023): 113405. </li> </ol> </div> <script type="text/javascript"> window.addEventListener('load',function(){ faqlist(4); }); </script> <h2 class="h2_tit page_tab_margin" id="tab6"> FAQ </h2> <ol class="ollist faqlist"> <li> <span class="sFAQTit">What is hybridoma sequencing?</span> <p> Hybridoma sequencing is a molecular technique used to determine the specific genetic sequence of the variable regions of both heavy and light chains in monoclonal antibodies produced by hybridoma cells. This method is essential for characterizing and replicating monoclonal antibodies for research and therapeutic purposes. </p> </li> <li> <span class="sFAQTit">Why is hybridoma sequencing important?</span> <p> Hybridoma sequencing is crucial for ensuring the specificity and efficacy of monoclonal antibodies. By determining the exact sequence of the antibodies, researchers can reproduce them reliably, study their properties in detail, and modify them for improved performance in diagnostic or therapeutic applications. </p> </li> <li> <span class="sFAQTit">What are the steps involved in hybridoma sequencing?</span> <p> The typical steps in hybridoma sequencing include: 1) Isolating RNA from hybridoma cells, 2) Converting RNA into cDNA using reverse transcription, 3) Amplifying the antibody gene segments using PCR, and 4) Sequencing the PCR products to identify the variable regions of the antibody genes. This process allows researchers to obtain the precise sequence data necessary for further analysis and application. </p> </li> <li> <span class="sFAQTit">Can hybridoma sequencing be used for any type of antibody?</span> <p> Hybridoma sequencing can be applied to any type of antibody produced by hybridoma cells. It is a versatile technique that can be used to sequence both monoclonal and polyclonal antibodies, although it is primarily used for monoclonal antibodies due to the nature of hybridoma technology, which produces a single type of antibody molecule. </p> </li> <li> <span class="sFAQTit">How accurate is hybridoma sequencing?</span> <p> Hybridoma sequencing is highly accurate when performed with modern sequencing technologies. It provides precise information about the variable regions of antibody genes, ensuring detailed and reliable characterization of monoclonal antibodies. The accuracy of the sequencing results depends on the quality of the sample preparation, the efficiency of the RNA extraction and PCR amplification, and the resolution of the sequencing platform used. </p> </li> <li> <span class="sFAQTit">What challenges might one encounter during hybridoma sequencing?</span> <p> Challenges in hybridoma sequencing can include inadequate RNA quality or quantity, which can compromise the cDNA synthesis; PCR amplification biases or errors, particularly in GC-rich or complex regions; and sequencing artifacts or errors. Overcoming these challenges often requires optimization of sample preparation protocols, careful design of PCR primers, and the use of high-fidelity enzymes and reliable sequencing platforms. </p> </li> <li> <span class="sFAQTit">How long does hybridoma sequencing typically take?</span> <p> The duration of hybridoma sequencing can vary depending on the specific protocols and equipment used, but it typically takes between a few days to a week. This time frame includes RNA extraction, cDNA synthesis, PCR amplification, and the actual sequencing process. Additional time may be required for data analysis and interpretation of the sequencing results. </p> </li> <li> <span class="sFAQTit">Can hybridoma sequencing be automated?</span> <p> Several aspects of hybridoma sequencing, such as PCR amplification and sequencing, can be automated to increase throughput and reduce human error. Automation is particularly useful in high-throughput environments where large numbers of monoclonal antibodies need to be sequenced. Automated systems can help streamline the workflow, from sample preparation to data analysis, enhancing the efficiency and reproducibility of the sequencing process. </p> </li> </ol> <h2 class="h2_tit page_tab_margin" id="tab7"> Resources </h2> <p> Use the resources in our library to help you understand your options and make critical decisions for your study. </p> <div class="tab_nav"> <span class="tab_choosebg">Videos</span><span>Podcasts</span><span>Infographic</span><span>Flyer</span><span>Case study</span><span>Articles</span> </div> <!---------Videos---------> <div class="tab_cont" style="display:block;"> <div class="row pagerow_pt"> <div class="col-md-4"> <div class="pagerow_pt_unit"> <img loading="lazy" src="../static/img/introduction-of-recombinant-antibody.jpg" alt="" /> </div> <a href="https://youtu.be/0-yZ9dTGg18" target="_blank">Introduction of Recombinant Antibody</a> </div> <div class="col-md-4"> <div class="pagerow_pt_unit"> <img loading="lazy" src="../static/img/immunogenicity-overview-of-therapeutic-biologics.jpg" alt="" /> </div> <a href="https://youtu.be/J430752_dTo" target="_blank">Immunogenicity overview of therapeutic biologics</a> </div> <div class="col-md-4"> <div class="pagerow_pt_unit"> <img loading="lazy" src="../static/img/video-development-of-monoclonal-antibodies-s.png" alt="" /> </div> <a href="https://youtu.be/NGqipRkxXiY" target="_blank">Antibody Library Screening - Creative Biolabs</a> </div> <div class="col-md-4"> <div class="pagerow_pt_unit"> <img loading="lazy" src="../static/img/antibody-based-cancer-therapies-creative-biolabs.jpg" alt="" /> </div> <a href="https://youtu.be/_Cqj5gphNug" target="_blank">Antibody based Cancer Therapies - Creative Biolabs</a> </div> <div class="col-md-4"> <div class="pagerow_pt_unit"> <img loading="lazy" src="../static/img/video-de-novo-antibody-sequencing-service.jpg" alt="" /> </div> <a href="https://youtu.be/lcdGTNKbaGQ" target="_blank"><em>De Novo</em> Antibody Sequencing Service</a> </div> <div class="col-md-4"> <div class="pagerow_pt_unit"> <img loading="lazy" src="../static/img/video-100-identification-of-leucine-and-isoleucine-in-de-novo-antibody-sequencing.jpg" alt="" /> </div> <a href="https://youtu.be/lcdGTNKbaGQ" target="_blank">100% Identification of Leucine and Isoleucine in <em>De Novo</em> Antibody Sequencing</a> </div> </div> </div> <!---------Podcasts---------> <div class="tab_cont"> <div class="therapy_ad"> <a href="therapeutic-antibody.html"> <img loading="lazy" src="../static/img/podcasts-ad.png" width="1700" height="500" srcset="../static/img/podcasts-ad.webp 1x,../static/img/podcasts-ad.webp 2x,../static/img/podcasts-ad.webp 3x" alt="In this brand new podcast series, we will show you everything you want to know about CAR-T therapy, including the mechanism, current applications, technology limitations, and potential strategies." /></a> </div> </div> <!---------Infographic---------> <div class="tab_cont"> <div class="row pagerow_pt"> <div class="col-md-4"> <div class="pagerow_pt_unit_2"> <img loading="lazy" src="/resource/images/A-Brief-Introduction-to-Antibodies-1.jpg" width="600" height="500" alt="" /> </div> <a href="https://go.creative-biolabs.com/a-brief-introduction-to-antibodies.html">A Brief Introduction to Antibodies</a> </div> <div class="col-md-4"> <div class="pagerow_pt_unit_2"> <img loading="lazy" src="/resource/images/Antibody-Library-Screening-Creative-Biolabs.jpg" width="600" height="500" alt="" /> </div> <a href="https://go.creative-biolabs.com/antibody-library-screening.html ">Antibody Library Screening</a> </div> <div class="col-md-4"> <div class="pagerow_pt_unit_2"> <img loading="lazy" src="/resource/images/De-Novo-Antibody-Seuqencing-Service-Creative-Biolabs.jpg" width="600" height="500" alt="" /> </div> <a href="https://go.creative-biolabs.com/de-novo-antibody-sequencing-service.html"><em>De Novo</em> Antibody Sequencing Service</a> </div> <div class="col-md-4"> <div class="pagerow_pt_unit_2"> <img loading="lazy" src="/resource/images/Antibody-Based-Therapies-1.jpg" width="600" height="500" alt="" /> </div> <a href="https://go.creative-biolabs.com/antibody-based-therapies.html">Antibody-Based Therapies</a> </div> </div> </div> <!---------Flyer---------> <div class="tab_cont"> <div class="row pagerow_pt"> <div class="col-md-4"> <div class="pagerow_pt_unit_2"> <img loading="lazy" src="/resource/images/Magic-Therapeutic-Antibody-Discovery-Platform-Flyer-Creative-Biolabs.jpg" width="600" height="500" alt="" /> </div> <a href="/resource/pdf/com/flyers/Magic-Therapeutic-Antibody-Discovery-Platform-Flyer-Creative-Biolabs.pdf">Magic<sup>TM</sup> Therapeutic Antibody Discovery Platform</a> </div> </div> </div> <!---------Case study---------> <div class="tab_cont"> <div class="row pagerow_pt"> <div class="col-md-4"> <div class="pagerow_pt_unit_2"> <img loading="lazy" src="/resource/images/Antibody-Development-Case-Study-com.jpg" width="530" height="500" alt="" /> </div> <a href="https://go.creative-biolabs.com/antibody-development-case-study.html">Antibody Development Case Study</a> </div> </div> </div> <!---------Articles---------> <div class="tab_cont"> <div class="row pagerow_pt"> <div class="col-md-4"> <div class="pagerow_pt_unit"> <img loading="lazy" src="../static/img/what-is-antibody-de-novo-sequencing-1.jpg" width="600" height="500" alt="" /> </div> <a href="/what-is-antibody-de-novo-sequencing.html">What Is Antibody <em>De Novo</em> Sequencing?</a> </div> <div class="col-md-4"> <div class="pagerow_pt_unit"> <img loading="lazy" src="../static/img/what-is-protein-mass-spectrometry-technology-1.jpg" width="600" height="500" alt="" /> </div> <a href="/what-is-protein-mass-spectrometry-technology.html">What Is Protein Mass Spectrometry Technology?</a> </div> <div class="col-md-4"> <div class="pagerow_pt_unit"> <img loading="lazy" src="../static/img/hybridoma-sequencing-technology-assisting-the-r-d-and-production-1.jpg" width="600" height="500" alt="" /> </div> <a href="/hybridoma-sequencing-technology-assisting-with-the-r-d-and-production-of-monoclonal-antibodies.html">Hybridoma Sequencing Technology - 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