Lactoferrin (LF) is a sialylated iron-binding glycoprotein, occurring in several biological secretions like milk, saliva, and seminal fluids and is a major component of a mammalian innate immune system. It plays multiple protective roles against large group of microorganisms and performs anti-inflammatory and anti-cancer activities. The concentration of serum LF in gilt (primiparous sow) and their piglets remains unknown. We determined serum LF concentration in gilts during gestation and lactation to that of 19-d-old piglets, including sow-fed and formula-fed piglets using enzyme-linked immunosorbent assay (ELISA). We found that the concentration of serum LF in gilts varied during gestation (0.77 ± 0.10 µg/mL) and lactation (0.62 ± 0.11 µg/mL). The mean concentration of serum LF in gilts (0.72 ± 0.06 µg/mL) was significantly higher than that of piglets (0.42 ± 0.07 µg/mL, P = 0.004). Additionally, a marginal significant difference (P =0.06) was observed for serum LF concentration in sow-fed piglets (0.42 ± 0.03 µg/mL) at 19 d old compared to that of formula-fed piglets (0.33 ± 0.04 µg/mL) at 37 d old. This study provides noble information regarding the serum LF concentration in the healthy gilts and piglets and thereby the data can be used as a standard reference point for future studies on the role of LF in pig reproduction.