Camel VHH Antibody Discovery Service

VHH antibodies (also known as nanobodies, single domain antibodies, or sdAbs) have been used in diagnostics, especially imaging, therapeutics (therapeutic antibodies, bispecific antibodies, CAR T, and ADCs), manufacturing, and research reagents. VHH antibodies are distinguished from other conventional antibody formats by their unique properties of size, solubility, intrinsic stability, easy tailoring into pluripotent constructs, recognition of uncommon or hidden epitopes, binding into cavities or active sites of enzyme targets, ease and speed of drug discovery, and ease of manufacture.

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 Library Discovery Service Background

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.

 

The single-domain nature of these antibodies provides unique advantages that set them apart for biotechnology applications. Some of these advantages include,

a) Stability at high temperatures & pH.

b) VHHs can recognize antigenic sites that are normally not recognized by conventional antibodies.

c) Their small molecular size helps with rapid tissue penetration and labeling applications, including crossing the blood-brain barrier.

d) Less expensive for large-scale production.

Camel VHH Antibody Library Discovery Service

 

 

Camel VHH Antibody Library Discovery Service Details

 

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

 

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 the VHH antibody supernatant

--10-15 expressing strains of the positive clone

Service Features of Camel VHH Antibody Library Discovery Service

--Provide immune library construction services and natural alpaca antibody library screening services

--A high-quality library and monoclonal antibody cell line can be obtained in approximately 16-20 weeks

--High library capacity: adequate library capacity 108-109/immune library, 109-1011/natural library

--Constructing high-affinity antibodies

--Personalized planning strategy

--Complete document traceability system and QC quality control file