Human Lactoferrin

1. Human Lactoferrin Lactoferrin is a glycoprotein secreted by mucosal membranes; it is a major protein in the secondary granules of leukocytes During intestinal inflammation, activated leukocytes infiltrate the mucosa and lumen, increasing the level of fecal lactoferrin Fecal lactoferrin is elevated in patients suffering from active inflammatory bowel disease (IBD) but not in persons with irritable bowel syndrome (IBS) or healthy persons

2. Lactoferrin levels in disease Ulcerative Colitis N=71 1200 800 400 0 Lactoferrin levels (ug/g wet weight) Crohn’s Disease N=78 Irritable Bowel Syndrome N=31 Healthy Persons N=56

3. Two new noninvasive tests to detect intestinal inflammation • IBD-CHEKTM – a qualitative ELISA for the clinical lab • IBD-QUIK CHEKTM – a qualitative lateral flow test for the clinical lab or physician’s office

4. IBD-CHEK TM Test • Qualitative ELISA for detecting elevated fecal lactoferrin as an indicator of intestinal inflammation • Utilizes a four-step procedure with results in 75 minutes • A valuable noninvasive diagnostic aid to distinguish active IBD from IBS • Available by direct order for hospital and reference labs

5. Performance characteristics of the IBD-CHEK TM Test N=236 Total IBD-CHEKTMIBD-CHEKTM Crohn’s disease Active 52 85% 15% Inactive 26 62% 39% Ulcerative colitis Active 40 88% 13% Inactive 31 52% 48% Active IBS 31 0% 100% Healthy persons 56 0% 100% (+) (-)

6. Clinical results with the IBD-CHEK TM • When distinguishing active IBD from irritable bowel syndrome and healthy persons, the IBD-CHEKTM test exhibited a sensitivity of 86% and specificity of 100% • When distinguishing active Crohn’s disease from irritable bowel syndrome and healthy persons, the IBD-CHEKTM test exhibited a sensitivity of 85% and specificity of 100%

7. IBD-QUIK CHEK TM Test • Lateral flow test for detecting elevated fecal lactoferrin as an indicator of intestinal inflammation • Helps to distinguish active IBD from IBS • Turnaround time in 10 minutes using a single specimen dilution • Suitable for use in the clinical lab or Physician’s office

8. Performance characteristics of the IBD-QUIK CHEKTM Test N=137 Total IBD-QUIK IBD-QUIK CHEKTM(+) CHEKTM (-) Crohn’s disease Active 58 100% 0% Inactive 35 6% 94% Ulcerative colitis Active 12 100% 0% Inactive 11 0% 100% Active IBS 17 0% 100% Healthy persons 27 0% 100%

9. Clinical results with the IBD-QUIK CHEKTM • When compared to the IBD-CHEKTM test for distinguishing active IBD from active irritable bowel syndrome and healthy persons, the IBD-QUIK CHEKTM test showed a sensitivity of 100% and specificity of 98% • When distinguishing active IBD from inactive IBD and active irritable bowel syndrome, the IBD-QUIK CHEKTM test showed a sensitivity of 100% and specificity of 96%

10. Fecal lactoferrin testing • The IBD-CHEKTM and IBD-QUIK CHEKTM tests are sensitive and specific in vitro diagnostic aids for distinguishing active IBD from IBS • Both assays offer a cost effective method for screening patients • IBS accounts for 2.4 to 3.4 million office visits annually; fecal lactoferrin testing can help distinguish this condition from more serious IBD

11. References 1. Dwarakanath, A. D., I. A. Finnie, C. M. Beesley, G. M. O’Dowd, J. Nash, H. H. Tsai, N. Parker, and J. Rhodes. 1997. Differential excretion of leucocyte granule components in inflammatory bowel disease: implications for pathogenesis. Clinical Science. 92: 307-313. • Guerrant, R. L., V. Araujo, E. Soares, K. Kotloff, A. Lima, W. Cooper, and A. Lee. 1992. Measurement of fecal lactoferrin as a marker of fecal leukocytes. J. Clin. Microbiol. 30:1238-1242. • Kane, S., Sanborn, W. Rufo, P., Zholudev, A., Boone, J., Lyerly, D., Camilleri, M., and S. Hanauer. 2001. Fecal lactoferrin is a sensitive and specific marker for chronic inflammatory bowel disease. Abstract of Digestive Disease Week / American Gastroenterology Association, Atlanta, Georgia. • Kayazawa, M., O. Saitoh, K. Kojima, K. Nakagawa, S. Tanaka, K. Tabata, R. Maysuse, K. Uchida, M. Hoshimoto, I. Hirata, and K. Katsu. 2002. Lactoferrin in whole gut lavage fluid as a marker for disease activity in inflammatory bowel disease: Comparison with other neutrophil-derived proteins. Am. J. Gastroenterol. 97:360-369. • Fine, K.D., F. Ogunji, J. George, M. Niehaus, and R. Guerrant. 1998. Utility of a rapid fecal latex agglutination test detecting the neutrophil protein, lactoferrin, for diagnosing inflammatory causes of chronic diarrhea. Am. J. Gastroenterol. 93:1300-1305. 6. Longstreth, G. F. , A. Wilson, K. Knight, J. Wong, C. Chiou, V. Barghout, F. Frech and J. Ofman. 2003. Irritable bowel syndrome, health care use, and costs: A U.S. managed care perspective. Am. J. Gastroenterol. 98:600-607. 7. Riley, L. W. 1995. Acute inflammatory diarrhea. In M. Blaser (ed.), P. Smith, J. Ravdin, H. Greenberg, and R. Guerrant, Infections of the Gastrointestinal Tract. Raven Press, New York, NY. 8. Sugi, K., Saithoh, O., Hirata, I., and K. Katsu. 1996. Fecal lactoferrin as a marker for disease activity in inflammatory bowel disease: Comparison with other neutrophil-derived proteins. Am. J. Gastroenterol. 91:927-934. 9. Uchida, K., R. Matsuse, S. Tomita, K. Sugi, O. Saitoh, and S. Ohshiba. 1994. Immunochemical detection of human lactoferrin in feces as a new marker for inflammatory gastrointestinal disorders and colon cancer. Clin. Biochem. 27:259-264.