How to make immunoglobulin after a tick bite? - briefly
After a tick exposure, collect the patient’s serum, isolate the specific anti‑tick IgG via affinity chromatography, and formulate it under sterile conditions for therapeutic use. Alternatively, immunize laboratory animals with tick antigens, generate monoclonal antibodies from hybridoma cells, and scale production in bioreactors.
How to make immunoglobulin after a tick bite? - in detail
After a tick attachment, the immune system recognizes antigens from the transmitted pathogen and initiates antibody production. Laboratory generation of specific immunoglobulin for therapeutic use follows a defined sequence.
First, confirm the presence of a tick‑borne infection through serologic or molecular testing. Identify the causative organism (e.g., Borrelia burgdorferi, Rickettsia spp.) and collect peripheral blood from a donor with a robust immune response, typically a convalescent patient.
Next, isolate peripheral blood mononuclear cells (PBMCs) and enrich for B lymphocytes using magnetic‑activated cell sorting or flow cytometry. Activate B cells with antigen‑specific stimulation (recombinant pathogen proteins, peptide epitopes) and cytokine cocktails (IL‑2, IL‑4, IL‑21) to promote differentiation into antibody‑secreting cells.
To create a renewable source, fuse activated B cells with an immortal myeloma line, generating hybridomas. Culture hybridomas in selective medium (HAT) and screen supernatants for target specificity by ELISA, western blot, or surface plasmon resonance. Clone positive hybridomas by limiting dilution to obtain monoclonal lines producing high‑affinity IgG.
Scale up production in bioreactors using serum‑free media. Harvest culture fluid, perform protein A/G chromatography to capture IgG, followed by ion‑exchange and size‑exclusion steps to remove aggregates and contaminants. Validate purity (>95 %), endotoxin levels (<0.1 EU/mL), and functional activity through neutralization assays.
Finally, formulate the purified antibody in a stabilizing buffer (histidine, sucrose), filter sterilize, and store at 2–8 °C. Release criteria include sterility, potency, and compliance with regulatory guidelines before clinical administration.
Key considerations:
- Early diagnosis improves donor antibody titers.
- Hybridoma technology yields consistent monoclonal products; alternatively, recombinant expression in CHO cells can be employed.
- Quality control must address specificity, affinity, and safety parameters.
The resulting immunoglobulin preparation can be administered prophylactically or therapeutically to patients at risk of severe tick‑borne disease, providing passive immunity while the host mounts its own response.