Prostate Cancer (PCa) is one of the leading medical issues faced by men worldwide and is the most prevalent cancer diagnosed in men in both Europe and the United States. The vast majority of current literature concerning PCa diagnosis concludes that better PCa markers are needed to reduce over-diagnosis of low risk disease and widespread over-treatment. The primary objective of this research entails the generation and characterisation of antiprostate specific antigen (PSA) isoform-specific recombinant antibody fragments. Selective screening of avian single chain antibody fragment (scFv) libraries was carried out for the isolation of high-affinity, anti-fPSA and anti-cPSA specific antibody fragments. These antibodies were kinetically evaluated using surface plasmon resonance-based instrumentation and incorporated into an enzyme-linked immunosorbent assay (ELISA) that can detect PSA in the ng/mL range. Anti-fPSA scFvB8 is a nanomolar affinity antibody fragment that contains highly conserved cysteine residues. High resolution X-ray crystallography of this antibody fragment and mutant variants was successfully carried out and revealed interesting structural attributes of avian antibody fragments. Glycan expression patterns change with the cellular modifications that accompany the onset of tumourigenesis. Hence, as a secondary objective, whole serum N-glycan profiling was carried out on 117 prostate cancer patient’s serum using a robotised, high-throughput analysis platform for glyco-profiling which utilises ultra-performance liquid chromatography (UPLC) to obtain high resolution separation of N-linked glycans released from serum. A specific glycomic signature identified from this analysis can distinguish between indolent and aggressive prostate cancer with high accuracy. The reagents generated and the results obtained from this body of work provide significant insight into antibody and glycomic-approaches that can potentially deliver the much sought after cancer-specific biomarker analysis for improved PCa diagnosis.