Snakebite envenoming is predominantly a life-threatening occupational disease caused by venom proteins in the bite of a venomous snake. The development of reliable rapid diagnostics is the need of the hour for timely snakebite diagnosis and the early administration of antivenom for treatment. Magnetic nanoparticles (MNPs) conjugated with antibodies have been extensively explored for the selective detection of biomarkers for diagnostic purposes. The purpose of the current study is to develop antivenom antibody-immobilized MNPs for the efficient immunodetection of snake venom proteins. Iron oxide magnetic nanoparticles were synthesized by the co-precipitation of Fe²⁺ and Fe³⁺ in sodium hydroxide (NaOH) solution. These MNPs were functionalized by sodium citrate solution. The synthesized citrate-capped MNPs with at least one carboxylic acid group on the surface were used for the immobilization of F(ab¢)₂ antibodies present in commercially available polyvalent antivenom. For the site-directed immobilization of antibodies, EDC-NHS ((1-ethyl-3-(3-dimethylaminopropyl) carbodiimide hydrochloride/N hydroxysuccinimide)) was used as linker. The antibody-immobilized and non-immobilized MNPs were characterized by X-ray diffraction and Fourier-transform infrared spectrometry analysis, and their size was estimated with the help of zeta potential. To validate the performance of antibody-conjugated MNPs in detecting snake venom proteins, a dot blot assay was performed using Russell’s viper venom protein antigens. The presence of snake venom-specific phospholipase A₂ (PLA₂) in human serum is an early indicator of snake bite envenomation and serves as a potential biomarker for the detection of free-flowing venom protein. Thus, antibody-immobilized MNPs were used as an immunoaffinity-based platform to capture a purified viper venom PLA₂. The present study demonstrated the successful conjugation of antibodies to the surface of magnetic nanoparticles and their ability to recognize viper venom proteins. Further, antibody-coated MNPs displayed the capability to effectively immunocapture a venom PLA₂ protein. Such antibody-coated nanoparticle-based immunodetection platforms will provide efficient enrichment and selective separation of snake venom-specific venom proteins from human serum. Additionally, immunoaffinity platforms based on nanoparticles for snake venom proteins with high sensitivity and high purity, and quick characterization will facilitate the early diagnosis of snakebite envenomation.