KMD Bioscience Co., Ltd.
Service Line：+86-400-621-6806
Address：FL-4, Building A5, International Enterprise Community, Tianjin, China
Email：info@kmdbioscience.com
Hundreds of alpaca VHH antibody library construction services have been successfully delivered every year. There is a growing number of related research. KMD Bioscience is experienced in the generation of alpaca VHH antibody and has been committed to this field for over 9 years. KMD Bioscience has established a comprehensive phage antibody display technology platform. Supported by this phage platform, KMD Bioscience can provide you with high-specificity and high-affinity alpaca VHH antibodies in a short period of time. More time for research, more chance of success.
Alpaca VHH Antibody Library Construction Background:
The alpaca is a member of the Camelidae family, which also includes llamas and camels. In addition to conventional IgG, these animals produce a functional single chained antibody that is capable of antigen binding sometimes referred to as VHH antibodies. These antibodies consist of only heavy chains and lack the light chains and functional CH1 and CH4 domains. These so-called heavy chain antibodies are being used to develop single-domain antibodies. The single domain nature of these antibodies provides unique advantages which set them apart for biotechnological applications.
Fig 1 Schematic representation of the peptide domains for camelid antibodies
Advantages of VHH Antibody: (1) stability at high temperatures and pH; (2) VHHs can recognize antigenic sites that are normally not recognized by conventional antibodies; (3) their small molecular size helps with rapid tissue penetration and labeling applications including crossing blood-brain barrier; (4) cheaper alternative for large scale production.
KMD Bioscience uses the pIII protein of M13 phage for VHH antibody (nanobody) surface display. Our scientists constructed a phage vector with Flag and 6×His tag double tags, and prepared TG1 and XL1-Blue host competent cells with 108 transfection efficiency, after 2-3 transfections and amplification, it can provide customers with a high-quality VHH antibody display library with an adequate library capacity of 108-109 and a packaging capacity of 1013-1016/ml. Up to 20-30 high-affinity phage strains can be obtained by three-round nanobody library screening by antigen-limited pressure screening. Through simple IPTG induction, a large number of antibody expression products can be obtained within 1-2 weeks to meet the research needs of customers, such as FACS and cell activity testing.
Service Process:
Service Procedure:
|
Steps |
Specification |
Timeline |
|
Step1: Alpaca Immunization |
* A healthy, full-aged Alpaca (28-36 months old) was immunized 4-6 times * Serum titers were measured by ELISA and collected peripheral blood |
8 weeks |
|
Step2: cDNA synthesis (Clients can provide cells) |
* Total RNA extraction * PCR amplification with Actin-specific primers to identify the quality of cDNA |
1 day |
|
Step3: VHH antibody Library construction |
* Primer design & synthesis * PCR amplification of variable region genes of heavy using cDNA as a template * Plasmid construction & transformation: after enzyme digestion, VHH and phagemid vector were ligated and transformed into TG1 host bacteria by an electric shock to construct antibody libraries * Identification: 20-50 clones were randomly selected, PCR identification, sequencing and analysis of antibody sequences |
3-4 weeks |
|
Step4: VHH antibody Library Screening (3 Rounds) |
* The antigenic limit pressure method is used for 3 rounds of panning
* Identification: 96 clones were randomly selected for ELISA titer determination * Up to 96 positive clones were sequenced, and the 10-15 clones were selected for prokaryotic expression transformation *10-15 positive clones were expressed to construct VHH antibody supernatant |
3-4 weeks |
Deliverables:
--Experimental report: including detailed construction procedures and representative sequence information
--VHH antibody library 2ml, >1⨯1013 /ml phage particles, Effective storage capacity>109
--10-15 positive clones were expressed to prepare VHH antibody supernatant
--10-15 expressing strains of positive clone
Service Highlights:
--Provide immune library construction services and natural alpaca antibody library screening services
--Short cycle, high-quality library and monoclonal antibody cell line can be obtained in about 16-20 weeks
--High library capacity: adequate library capacity 108-109/immune library, 109-1011/natural library
--High-affinity antibody construction
--Personalized panning strategy
--Complete document traceability system and QC quality control file
How to Order?
If you have any questions regarding our services or products, please feel free to contact us by E-mail: info@kmdbioscience.com or Tel: +86-400-621-6806
