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Nanobodies, also known as variable heavy chain (VHH) antibodies, represent the smallest fully functional antigen-binding fragments. With a molecular weight of only ~15 kDa—about one-tenth that of conventional antibodies—they preserve exceptional affinity and specificity, making them invaluable tools for precision medicine, disease diagnostics and therapeutic development. In Mycobacterium tuberculosis research, nanobodies combine the advantages of traditional antibodies and small-molecule drugs through their strong tissue penetration and remarkable stability, offering broad prospects for diagnosis and treatment and introducing a novel strategy to address drug-resistant tuberculosis.
Fig. 1 General structure of antibody fragments.
Nanobodies can be generated from immune, naïve, or synthetic libraries. In the case of immune libraries, camelids such as llamas or alpacas are immunized with the target antigen (or transgenic mice are used when antigen availability is limited). Peripheral blood is then collected, and lymphocytes are isolated for mRNA extraction and cDNA synthesis. The VHH (heavy-chain variable) domains are amplified by PCR, cloned into display vectors, and transformed into bacterial, phage, or yeast systems, creating a large and diverse pool of nanobody candidates. Following at least three rounds of screening—such as liquid-phase, solid-phase, and cell-based screening—high-affinity clones are identified.
Top-performing candidates are subsequently expressed in high-efficiency recombinant systems to produce soluble VHH or VHH-Fc fusion nanobodies at scale. Each purified nanobody undergoes thorough characterization for antigen-binding specificity, stability, and other critical performance attributes, ensuring consistent quality for downstream diagnostic or therapeutic applications.
Fig. 2 A schematic diagram of the different methods of generating Nanobodies
Creative Diagnostics offers nanobodies targeting more than ten critical Mycobacterium tuberculosis antigens—including TB15.3, Mpt64, Mpt83, Ag85A, Ag85B and Ag85C. Several of these are available as matched pairs, making them ideal for sandwich ELISA development or dual-epitope detection formats and providing assay developers with a strong foundation for creating high-sensitivity, low-background tests.
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