Truncated PAP2 and methods of making and using same
View Patent ↗A method of treating pancreatitis is provided, including the steps of: providing a mammal having pancreatitis; and administering a therapeutically effective amount of a truncated N-terminal PAP2; and making of an antibody specifically directed to detect PAP2.
1. A truncated pancreatitis associated protein of the sequence listing
(SEQ ID NO: 1)
LVTTLKSWFQADLACQKRPSGHLVSILSGGEASFVSSLVTGRVNNNQDIW
IWLHDPTMGQQPNGGGWEWSNSDVLNYLNWDGDPSSTVNRGNCGSLTATS
EFLKWGDHHCDVELPFVCKFKQ.
2. The truncated pancreatitis associated protein of claim 1 , wherein said protein is in substantially purified form.
3. A peptide sequence usable to make anti-PAP2 antibody of the sequence TMGQQPNGGGWEWSNSDVLNYLNWDGDPSST (SEQ ID. NO. 25).
4. A method of treating pancreatitis, comprising:
providing a mammal having pancreatitis; and
administering a therapeutically effective amount of truncated pancreatitis associated protein of the sequence listing
(SEQ ID NO: 1)
LVTTLKSWFQADLACQKRPSGHLVSILSGGEASFVSSLVTGRVNNNQDIWI
WLHDPTMGQQPNGGGWEWSNSDVLNYLNWDGDPSSTVNRGNCGSLTATSEF
LKWGDHHCDVELPFVCKFKQ.
5. The method of claim 4 , further comprising the step of correlating a dose with a result.
6. The method of claim 5 , wherein said result is selected from the group consisting of clinical assessment of acute abdomen, white count, and elevated amylase and lipase levels.
7. The method of claim 4 , further comprising the step of observing the expression of NF-κB.
8. The method of claim 7 , further wherein observing is measured by a method selected from the group consisting of ELISA, a western blot, or a PCR.
9. A method of providing an immunomodulatory effect in a mammal with pancreatitis, comprising:
administering to the mammal in need thereof a therapeutically effective amount of a truncated pancreatitis associated protein of the sequence listing
(SEQ ID NO: 1)
LVTTLKSWFQADLACQKRPSGHLVSILSGGEASFVSSLVTGRVNNNQDIWI
WLHDPTMGQQPNGGGWEWSNSDVLNYLNWDGDPSSTVNRGNCGSLTATSEF
LKWGDHHCDVELPFVCKFKQ.
10. A method of making recombinant truncated pancreatitis-associated protein of the sequence listing
(SEQ ID NO: 1)
LVTTLKSWFQADLACQKRPSGHLVSILSGGEASFVSSLVTGRVNNNQDIWI
WLHDPTMGQQPNGGGWEWSNSDVLNYLNWDGDPSSTVNRGNCGSLTATSEF
LKWGDHHCDVELPFVCKFKQ,
comprising the steps of:
Inserting a nucleic acid encoding SEQ ID NO:1 in-frame into a pET24a bacterial expression vector;
growing a plurality of positive clones transformed into a plurality of bacterium to a density in a medium;
centrifuging and resuspending the bacterium in a resuspension buffer;
sonicating the bacterium with a protease inhibitor to lyse the cells;
centrifuging and washing the bacterial cell lysate pellet in a first buffer;
solubilizing said bacterial cell lysate pellet in a second buffer containing 6 M urea.