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Specific to Osteoclasts (Bone resorption)Specific to Osteoclasts (Bone resorption)

Sensitive to change in born metabolism Sensitive to change in born metabolism

Small day-to-day and diurnal variation Small day-to-day and diurnal variation

Measurable in blood specimenMeasurable in blood specimen

Unaffected by food-intakeUnaffected by food-intake

Unaffected by renal dysfunctionUnaffected by renal dysfunction

Measurable active TRACP-5b enzyme Measurable active TRACP-5b enzyme

Bone Resorption Marker

Measurement kit for tartrate-resistant acid phosphatase 5b

Reference values of TRACP-5bReference values

Reference values of TRACP-5b for female (YAM: young adult mean) were determined as 120 – 420 mU/dL based on the range of the mean ± 1.96 SD, established in healthy premenopausal women. For every women, whether premenopausal or postmenopausal, the reference values are applied. The cut-off value for a high future fracture risk is regarded as 420 mU/dL.

M a l eFemale (YAM value)

170 - 590 mU/dL120 - 420 mU/dL

(Nishizawa, Y., et al., Jpn J Med Pharm Sci 54:709, 2005)

(coloring)

wash &remove

enzyme reaction antigen-antibody reaction antibody plate

: TRACP-5a : TRACP-5b fragment (inactive)

: TRACP-5b (active): microwell plate

: anti-TRACP-5b antibody

: anti-TRACP-5b fragment antibody

: substrate (CNPP)

: product (CNP)

OSTEOLINKS TRAP-5b assay design and procedureOSTEOLINKS TRAP-5b is a novel fragments absorbed immunocapture enzymatic assay (FAICEA) kit. This assay system employs two unique monoclonal antibodies, one is specific to active TRACP-5b and the other is specific to inactive TRACP-5b fragments. The system enables highly specific measurement to TRACP-5b activity with wide dynamic range.

Basic performance of OSTEOLINKS TRAP-5bSample type serum, heparin-plasma

Sample stability [Serum] Room temp.(25ºC): 8 hours, 2 - 8ºC: 2 days, -20 - -30ºC: 1 month, -80ºC: 24 months, Maximum 3 freeze-thaw cycles [Whole blood] Room temp.(25ºC) or 2 - 8ºC: 12 hours Dietary variation TRACP-5b activity in blood has minimal response to food. There is no need for fasting before sampling.

Renal dysfunction effect TRACP-5b is degraded and inactivated during circulation. Osteolinks TRAP-5b measures enzymatically active TRACP-5b levels in blood, and renal dysfunction does not affect the levels.

OSTEOLINKS TRAP-5b productsAssay method EIA method (FAICEA : Fragments absorbed immunocapture enzymatic assay)

Packaging 96 assays Storage condition 2 - 10ºC

Shelf life 18 months after manufacturing

TRACP-5b (tartrate-resistant acid phosphatase 5b isozyme) is a 35-37 kDa glycoprotein. TRACP-5b is closely associated with osteoclast activity, and is secreted into the blood. Serum TRACP-5b measurement is valuable as a specific bone resorption marker. OSTEOLINKS TRAP-5b is a novel fragments absorbed immunocapture enzymatic assay (FAICEA) kit. The system enables highly specific measurement to TRACP-5b activity with wide dynamic range, without cross-reaction with TRACP-5a derived from macrophages.

For in Vitro Diagnostic Use Approved No. 22000AMX00076000

OSTEOLINKSTM TRAP-5bOSTEOLINKSTM TRAP-5b

TRA-008-1/2106GRA

https://www.sbbio.co.jp/

Clinical usefulness of OSTEOLINKS TRAP-5b for therapeutic monitoring in osteoporosisSerum TRACP-5b markedly decreased in the first month of the treatment, then the reduction continued until 3rd and 6th month (Figure 1). In the 36 cases whose LBMD had effectively increased in the first 6th month, serum TRACP-5b was significantly decreased in excess of MSC in 34 cases at the 1st month and 35 cases at the 3rd month (Figure 2).

Figure 1 Changes in the bone metabolic markers after bisphosphonate treatment for osteoporosis patients

Methods: Postmenopausal osteoporosis patients (n=57) were treated with bisphosphonates, and bone metabolic markers were measured at baseline and after 1, 3 and 6 months. Reduction rates, i.e., percentage changes from baseline, were indicated for each marker (mean ± SD).

(Miki, T. et al., Jpn J Med Pharm Sci 55: 443, 2006)

Figure 2 Rates of cases with change in bone metabolic markers exceeded the MSC in the 36 patients who had improved in their LBMD with bisphosophonate treatment. Methods: 57 postmenopausal osteoporosis patients were treated with bisphosphonates for 6 months, and 36 cases had improved with 3% or more increase from the baseline in LBMD. For the 36 cases, the rates of cases whose bone metabolism markers had changed in excess of the MSC at 1 and 3 months were examined retrospectively.

(Miki, T. et al., Jpn J Med Pharm Sci 55: 443, 2006)

Serum TRACP-5b Serum NTX Urine NTX Urine CTX Urine DPD Serum BAP

1 month 6 months3 months 1 month 3 monthsTreatment time Treatment time

Perc

ent c

hang

e fro

m b

asel

ine

Rates

of ca

ses w

ith ch

ange

in th

e mark

er ex

ceed

ed th

e MSC

in th

e 36 p

atien

ts wh

ose L

BMD

had e

ffecti

vely

increa

sed

--- : MSC for each marker

High detectability with small MSC (Minimum Significant Change)

Items SerumTRACP-5b

UrineNTX

UrineDPD

SerumNTX

UrineCTX

SerumBAP

Day-to-dayvariation

MSC

8.1% 29.9%32.7%17.4%17.5% 6.4%

16.2% 34.9% 34.8% 65.4% 59.9% 12.9%

Table 1 Minimum significant Change (MSC) for bone metabolism markers Methods:Minimum significant changes were calculated from the day-to-day variation data of each bone metabolism marker in Figure 3.

(Mochizuki, Y. et al., Jpn J Med Pharm Sci 54: 895, 2005)

Minimum Significant Change (MSC)MSC is one criterion for evaluating the effectiveness of the treatment for osteoporosis and other bone diseases by bone metabolism markers. The MSC is defined as twice the value of day-to-day variation, considering physiological and methodological variation of the marker. Therefore, to change in excess of this criterion means actual change caused by the therapeutic intervention. Using a marker with smaller MSC for evaluation enables to detect slighter change in the bone metabolism.

Characteristics of OSTEOLINKS TRAP-5b Small day-to-day and diurnal variations (High reproducibility) Day-to-day variation of serum TRACP-5b was much smaller than that of the other bone resorption markers. (Figure 3)Also, diurnal variation of serum TRACP-5b was smallest in the tested bone resorption markers. (Figure 4)

Serum TRACP-5b Serum NTX

Serum BAP

Urine DPD

Urine NTX Urine CTX

Perc

enta

ge to

ave

rage

Perc

enta

ge to

ave

rage

Sampling number Sampling numberSampling number

Figure 3 Examination of day-to-day variations of bone metabolism markers Methods: Bone metabolism markers were measured in 8 specimens from every 10 postmenopausal women. The specimens (blood and urine) were obtained from 9:00 to 11:00 am of 8 separate days at intervals of 4 days or more. The data are expressed as relative values, with the average of each subject being 100 percent.

(Mochizuki, Y., et al., Jpn J Med Pharm Sci 54: 895, 2005)

Serum TRACP-5b Serum NTX

Serum BAP

Urine DPD

Urine NTX Urine CTX

Morning Afternoon Evening Morning Afternoon EveningMorning Afternoon Evening

Morning Afternoon EveningMorning Afternoon EveningMorning Afternoon Evening

Sampling time Sampling time Sampling time

Perc

enta

ge to

ave

rage

Perc

enta

ge to

ave

rage

Figure 4 Examination of diurnal variations of bone metabolism markers Methods Bone metabolism markers were measured in 3 specimens from every 6 postmenopausal women. The specimens (blood and urine) were obtained in the morning (9:00-11:00am), afternoon (1:00-2:00pm), and evening (4:00-6:00pm) a single day. The data are expressed as relative values, with the average of each subject being 100 percent.

(Mochizuki, Y., et al., Jpn J Med Pharm Sci 54: 895, 2005)

Characteristics of OSTEOLINKS TRAP-5b