Design Goal : NGAL: A Rapid Biomarker for Acute Kidney InjuryTeam 1: Vinoudini Boj, Farhan Bukhari, Daniel Burke, Cijian Feng, James Y. Hasegawa, Alex Lee, Jessica Potts, Stephanie Wong

ALS 320 Design Project , Fall 2010, Keck Graduate Institute of Applied Life Science, Claremont, CA

Competitor Product Analysis

Clinical Utility

Market / Competitor Analysis

Executive Summary

NGAL is a 25-kDa extracellular lipocalin protein.

Massive up-regulation of NGAL following kidney damage and during

bacterial infection and inflammation (1, 2).

NGAL is released from proximal renal tubules soon after kidney damage

Dramatic increase of NGAL levels in urine and serum samples can be

detected in 2 hours (3, 4).

(a) (b)

Figure 1. (a) The NGAL level of urine samples increased dramatically in patients who later

develop AKI. (b) Receiver-operating characteristic curves demonstrated that the

measurement of urine NGAL level 2 hours after surgery is a sensitive and specific

diagnostic tool for AKI diagnosis (5).

Measuring NGAL level before surgery was found to enable identification of

kidney distress, which greatly increases the incidence of AKI post-

operatively (6).

Other useful AKI biomarkers:

Interleukin-18 - level elevated 4-6 hours after cardiac surgery

Cystatin C - level elevated within the first 12 hours and were sustained 24

hours post-surgery (7, 8)

1. J. Am. Soc. Nephrol. (2003) 14, 2534-2543. 2. Scand. J. Clin. Lab. Invest. (1995) 55, 125-131. 3. Am. J. Nephrol.

(2004) 24, 307-315. 4. J. Clin. Invest. (2005) 115, 610-621. 5. Lancet (2005) 365, 1231-1238. 6. Critical Care. 11,

R127. 7. Am. J. Transplant. (2006) 6, 1639-1645. 8. Clin. J. Am. Soc. Nephrol. (2010) 5, 1552-1557,.,

Acute Kidney Injury (AKI) is a critical condition afflicting 5% of hospital

admissions and up to 50% of intensive care unit (ICU) patients, often with fatal

outcomes. While measurement of the conventional biomarker, creatinine,

detects AKI 2-3 days after surgery, Neutrophil Gelatinase-Associated Lipocalin

(NGAL) detects AKI with high sensitivity and specificity in two hours. This time

difference provides a valuable treatment opportunity for successful prevention

of further kidney injury. In this review, we analyze NGAL as a new AKI

biomarker, evaluate current NGAL diagnostic devices, and assess the market

potential for a point-of-care NGAL assay.

Current assays and devices for measuring NGAL include the Abbott Architect

Analyzer, BioPorto NGAL Rapid ELISA Kit, and Biosite Triage NGAL Test.

Devices and technologies in development that could be used to measure NGAL

include Abbott i-STAT, and Proxim DX’s electronic sensor technology.

A NGAL point-of-care device that provides accurate, reliable results within two

hours of injury would meet the needs of clinicians in the critical care setting.

Use Environment Analysis and Use Case

Market size - $140 million to $371.6 million

Anywhere from 550,000 cases of AKI to potentially 2 to 3 million cases per

year in the US (9).

Mortality rate of 21.3% for AKI vs 2.3% for all hospital admissions (10)

AKI is caused by many common illnesses / conditions / medications

inaccurate prevalence

Value Proposition - The average postoperative cost is roughly $19,211 higher

for AKI patients than non-AKI patients(11).

9. Cli.n J. Am. Soc. Nephrol. (2005) 1, 6. 10. Clin. J. Am. Soc. Nephrol. (2008) 3, 481-490. 11. Nephrol. Dial.

Transplant. (2008) 23, 1970-1974

Point-of-care in hospital ICUs and other critical care units

Portable

Straightforward and simple administration

Accurate, unambiguous and widely accessible results

Cost-effective

Product Vision Statement and Unmet User Need

Current AKI biomarker creatinine takes 48-72 hours for the levels to be

sufficiently elevated.

Faster AKI diagnosis is needed to improve clinical outcomes

Our inexpensive, easy-to-use, portable, point-of-care (POC) NGAL device

based on bio-sensor technology will revolutionize AKI diagnosis by

providing fast and reliable data.

Competitor analysis:

NGAL test is not FDA-approved as a diagnostic biomarker of AKI in the U.S.

Abbott and Biosite are developing NGAL tests for their IVDs, Abbott

Architect and Biosite Triage.

BioPorto Diagnostics (Denmark) has an NGAL ELISA kit available in certain

markets outside the U.S.

Competing Biomarkers:

No IL-18 IVDs developed yet

Seven cystatin C IVDs in the market, including Dade Behring, Siemens, and

Diazyme Laboratories (12)

Patent Issues: Cincinnati Children’s Hospital (CCH), Phadia, BioPorto are

tangled in legal actions in the U.S. and Europe for sole rights to NGAL as a

biomarker for AKI. (13, 14)

12. U S food and drug administration home page http://www.fda.gov/. 13. BioPorto Annual Report 2009 . 14.

Interim Financial Report for Q2 2010 for the BioPorto Company.

Abbott Architect

• Chemiluminescent Magnetic Immunoassay

• Sandwich ELISA on magnetic beads

• Detection Ab linked to Acridium Ester

• Pre-trigger solution disrupts Analyte-AB interactionDetection Ab back in solution

• Beads removed

• Trigger reagent hydrolyzes Acridiumester to emit light

• Urine sample

• 30 minutes

• Expensive

• Core Lab

BioPorto Rapid ELISA Kit

• 96 well Plate

• Horseradish Peroxidase, TMB

• Absorbance measured at 450 nm

• Urine or plasma sample

• Detect urine range 110 ng/mL to 40,000 ng/mL

• 45 minutes

• Core Lab

Biosite Triage

• Lateral Flow-like Sandwich Immunoassay

• Filter separates plasma

• Capillary action moves sample to interact with Detection Ab

• Fluorescence Energy Transfer Particles

• Sample moved to zones of capture Ab

• Excited Particles Emit Photons

• Plasma sample

• Detection range: 60 ng/mL to 1,300 ng/mL

• 15 minutes

• POC

Abbott i-STAT

• Electrochemical Assay

• Sandwich

• Alkaline Phosphatase

• Blood sample

• Thin-film electrodes

• Automatic calibration for every sample

• Data sent to central database

• Minutes

• POC

Proxim DX

• Sandwich ELISA with Amperometric Electrochemical Detection

• Screen printed carbon electrodes

• Thick film microfabrication

• Instantaneous product formation measurement

• Inexpensive