Camel VHH Antibody Discovery Service

Introduction Of Camel VHH Antibody Discovery Service

Camel, as all members of the camelid family, produce antibodies with an original scaffold only made of heavy chains (IgG2 and IgG3), thus promoting binding to structural motifs unreachable before in protein targets, such as viral and enzymatic proteins. Due to the VHH fragments and smaller size, camel heavy-chain antibodies can transform into bacterial cells easier, which is ideal for research purposes. With further modification, the VHH fragments can also be developed as single-domain antibodies. Because engineering, selecting and producing these recombinant camel antibodies as single-domain antibodies (sdAbs), also called nanobodies for their shorter size, require outstanding competence and experience in recombinant antibody molecular and cell biology, few research teams in the world can deliver functional sdAbs for immunodiagnostic and immunotherapy.

Camel VHH Antibody Discovery Service Workflow

Steps	Specification	Timeline
Step1: Camel Immunization	--A healthy, full-aged Camel (28-36 months old) was immunized 4-7 times --Serum titers were measured by ELISA and collected peripheral blood	6-8 weeks
Step2: cDNA synthesis (Clients can provide cells)	-Total RNA extraction --RT-PCR --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
VHH-Fc Antibody Production (optional)	--Gene synthesis, subcloning, plasmid preparation --Transient expression and purification --QC analysis	2 weeks
Affinity Ranking (optional)	--Affinity measured by Biacore 8K	1 weeks

What We Can Offer?

Alpha Lifetech Inc. is a well-recognized leader in the field of single-domain antibody discovery. Alpha Lifetech Inc. has developed proprietary procedures for nanobody discovery. Our optimized library construction and screening processes with phage display technology guarantee successful isolation of high-affinity camel nanobodies in a short time. We have successfully produced multiple positive clones for a variety of antigens, ranging from large protein molecules (>400kD) to small haptens (small chemical molecules of ~200 daltons), many of the binders have sub-nanomolar affinities measured by ELISA.

Camel VHH Antibody Discovery Service Process

Alpha Lifetech can provide VHH antibody discovery service, including VHH antibody library construction service and VHH antibody library screening service. Deliverables of Alpha Lifetech: