Prolactin Chicken, PEG
Greater than 95.0% as determined by gel filtration analysis.
Description
Prolactin Chicken Recombinant Pegylated produced in E.Coli is a single, non-glycosylated polypeptide chain containing 199 amino acids + an additional Ala at n-terminal. Pegylated Prolactin is mono-pegylated having a molecular mass of ~ 39 kDa, however under non-denaturing conditions it behaves as 220 kDa protein due to its increased hydrodynamic volume.
The Pegylated Prolactin protein is purified by proprietary chromatographic techniques.
Product Specs
Prolactin, a neuroendocrine hormone primarily produced by the pituitary gland, is also synthesized in other cells like those found in the placenta, brain, and uterus. Its secretion is triggered by various stimuli such as eating, nursing, mating, estrogen treatment, and ovulation. While prolactin's primary function is to stimulate and maintain lactation, it also plays a role in breast cancer development, regulation of reproductive functions, and immunoregulation.
Recombinant Pegylated Chicken Prolactin, produced in E. coli, is a single, non-glycosylated polypeptide chain consisting of 199 amino acids and an additional alanine at the N-terminal. This pegylated prolactin is mono-pegylated with a molecular mass of approximately 39 kDa. However, under non-denaturing conditions, it exhibits a behavior similar to a 220 kDa protein due to its increased hydrodynamic volume.
The purification of the pegylated prolactin protein is achieved through proprietary chromatographic techniques.
The protein was lyophilized from a concentrated solution (1 mg/ml) containing 0.02% sodium bicarbonate (NaHCO3).
To reconstitute the lyophilized prolactin, it is recommended to dissolve it in sterile 0.4% sodium bicarbonate (NaHCO3) at pH 8.5, to a concentration of at least 100 µg/ml. This solution can then be further diluted in other aqueous solutions as needed.
Lyophilized prolactin, while stable at room temperature for up to 3 weeks, should ideally be stored desiccated at temperatures below -18°C. After reconstitution, prolactin should be stored at 4°C for 2-7 days. For long-term storage, freezing at -18°C is recommended.
For enhanced stability during long-term storage, adding a carrier protein like 0.1% HSA (Human Serum Albumin) or BSA (Bovine Serum Albumin) is recommended.
Avoid repeated freeze-thaw cycles to maintain protein integrity.
The purity is determined to be greater than 95% based on gel filtration analysis.
The biological activity of Pegylated Chicken Prolactin was assessed in vitro through its ability to induce proliferation in Nb2 cells and Baf/3 cells stably transfected with Chicken Prolactin receptors. While its activity is lower than non-pegylated Chicken Prolactin in vitro, it is expected to exhibit higher in vivo activity due to its prolonged circulation time resulting from pegylation.
Product Science Overview
Recombinant chicken prolactin (chPRL) is produced using Escherichia coli (E. coli) as the expression system . The protein is expressed as a single, non-glycosylated polypeptide chain containing 199 amino acids, with an additional alanine at the N-terminal . The recombinant protein is then purified to homogeneity.
PEGylation refers to the process of attaching polyethylene glycol (PEG) chains to a protein or peptide. This modification can enhance the pharmacokinetic properties of the protein, such as increasing its stability and prolonging its circulation time in the bloodstream . For chicken prolactin, PEGylation results in a mono-PEGylated protein (PEG-chPRL) with a molecular mass of approximately 39 kDa .
The biological activity of PEG-chPRL has been evaluated through its interaction with the human prolactin receptor extracellular domain (hPRLR-ECD) and its ability to stimulate PRLR-mediated proliferation in Nb2-11C cells . Although PEG-chPRL exhibits lower activity in vitro compared to non-PEGylated chPRL, it has a significantly prolonged circulation time, remaining in the bloodstream for 16 hours longer than the non-PEGylated form .
The PEGylation of chPRL leads to favorable pharmacokinetic alterations, allowing for reduced dosing frequency due to its prolonged persistence in the circulation . In vivo experiments have shown that PEG-chPRL injections in chickens result in more profound effects on corticosteroid levels in the blood compared to non-PEGylated chPRL .
The development of PEG-chPRL opens new avenues for poultry research, particularly in understanding the hormonal regulation of reproductive behaviors and stress responses. The improved pharmacokinetic properties of PEG-chPRL make it a valuable tool for in vivo studies, potentially reducing the frequency of dosing and improving the overall efficacy of the experiments .
In conclusion, the PEGylation of recombinant chicken prolactin represents a significant advancement in poultry research, offering enhanced stability and prolonged activity. Future research will likely focus on further optimizing the PEGylation process and exploring its applications in various physiological studies.
