Aim This study aimed to demonstrate biosimilarity between Lupin’s Pegfilgrastim versus Neulasta® with respect to pharmacokinetic (PK), pharmacodynamic (PD), immunogenicity, and safety profiles. Methods This double-blind, randomised, single-dose, two-sequence, two-treatment, two-period, crossover, comparative PK and PD study enrolled healthy subjects aged 18 to 45 years. The test product was Lupin’s Pegfilgrastim and the reference product was U.S.-licensed Neulasta®. Blood samples were collected at different time points for assessment of PK, PD, and immunogenicity. Primary objectives included comparing the relative bioavailability and the pharmacodynamic effect of the two products. Secondary objectives included assessing immunogenicity, safety and tolerability. Biosimilarity was concluded if the 90% confidence interval (CI) of the ratio of the geometric least-squares mean (LSM) for PK and PD parameters were within 80 to 125%. Results The geometric mean ratios of Lupin’s Pegfilgrastim and Neulasta® for primary PK parameters were Cmax [94.37 (89.53 to 99.47%)], AUC0-t [94.76 (89.83 to 99.95%)] and AUC0-∞ [94.82 (89.90 to 100.02%)], and for primary PD parameters were ANC_Cmax [100.59 (98.96-102.25%)] and ANC_AUEC0-tlast [100.51 (99.18,101.85%)]. The corresponding 90% CIs were within the pre-defined equivalence limit of 80% and 125%. The incidence of ADA formation was similar between patients receiving Lupin’s Pegfilgrastim and Neulasta®. No new safety concerns were observed. Conclusion This study establishes biosimilarity of Lupin’s Pegfilgrastim and Neulasta® with respect to PK and PD profiles. No clinically meaningful differences were observed between the biosimilar and the reference product in terms of immunogenicity, safety, and tolerability profiles.