Kanto Diagnostics

Characteristics
The measurement principle is Immunologicalinhibition method.
By the double kinetic mode, it can avoid the influenceof adenylate kinase by hemolysis.

Kanto Diagnostics
Principle②M B+M M

M M
CK-M inhibitory
antibody + substrate

M B①
Isozyme in serum
Creatinine kinase is composed ofdimer of M (muscle type) subunit andB (brain type) subunit.
It's widely known that there are 3 typeisozyme of MM, MB, BB.
But basically, BB does not exist in theblood.

Creatine phosphate + ADP
ATP + D-Glucose
D-Glucose-6-phosphate + NADP

M B

M subunit is inhibited by CK-M inhibitory
antibody. There are inhibition pattern offollowing 2 types.
① Steric hindrance
When the antibody binds to M subunit, thesubstrate can not reach the active site of theM subunit.
② Allosteric effect
Structural change will occur by binding
antibodies to any antigenic determinant of Msubunit.

CK-B

Calculate the CK-MB activity by
doubling the remaining B
subunit activity.

Creatine + ATPHK

ADP + D-Glucose-6-phosphate

G-6-PD

D-Gluconate-6-phosphate + NADPH + H+

Macro CKMacro CK TypeMacro CK is classified type 1 and type 2.
These macro CK are measured by total CK activity
and CK-MB activity.
Since CK-M inhibition antibody does not inhibit
macro CK, CK-MB will be abnormal high value in thecase that macro CK is high concentration.

Method of determining
CK-MB/CK > 25%

Type １

B B
Type 2mCKmCKmCKmCKmCKmCKmCK
mCK

Electrophoresis
Type1 is occupies a large
percentage of those in which
the immune globulin and CKBB is bound.
And Fab of lgG and CK-BB isbonded in a ratio of 2-to-1.

Type2 is octamer thatmitochondria CK(mCK) arepolymerized.

Heat inactivation test
（56℃, 10 minutes）** For example

CK Residual activity ≦50%

Not macro CKCK Residual activity ＞
50%

Macro CK
suspected

Kanto Diagnostics
Inhibitory capacity

Influence tests

80604020

製造直後capacity(0阻害率 month)
Inhibitory
Inhibitory
capacity(10months)
製造後10ヶ月
阻害率
Residualmonth)
製造直後activity(0残存活性
Residualactivity(10残存活性months)
製造後10ヶ月後

10003500

7000

10500

10509501514000-52500 5000 7500 10000
CK-MM活性(U/L)
CK-MM (U/L)

0CK－MM(U/L)
(U/L)
CK-MM

Stability
Shelf lifestability
製造後安定性

Open stability
開封後
安定性

250

275

200250

生理食塩水
Saline150225

コントロール血清
Control serum

10020050175
150

100806004681004Days
開封日数Linearly

Within-run precision

7000
6000

Serum
N
MEAN
MAX
MIN
RANGE
S.D.
C.V.

5000
4000
3000
2000
1000
002468300 400500

BIL-C
BIL-C
120
10080
6005101520
Concentration(mg/100mL)
BIL-C 濃度 (mg/100ml)
BIL-F
BIL-F

1401201008060058 12 16 20 24 28 32

製造後月数Months

12031.8333120.639
2.009

220257.7259
25630.7450.289

10Dilution
series (n/10)
希釈段階(n/10)*）Please dilute the samples in the case that total CK is more thaninhibitory capacity even if CK-MB is in the range of linearly.

CK-MB活性(U/L)
activity(U/L)
CK-MB

2100 200

14000(U/L)
activity(U/L)
CK-MB
CK-MB活性120101520BIL-F 濃度 (mg/100ml)
Concentration(mg/100mL)

乳びLipemic
14012010080600500

1000

1500

Concentration(Formazine turbidity)
乳び 濃度 (ホルマジン濁度)(U/L)
activityCK-MB
CK-MB活性(U/L)(U/L)
activity(U/L)
CK-MB
CK-MB活性300

ヘモグロビン
Hemolysis

140

Concentration
ヘモグロビン濃度(mg/100mL)(mg/100ml)

CK-MB活性(U/L)
activity(U/L)
CK-MB

Company
A
A社
免疫阻害法
CicaLiquid CK-MB
シカリキッド
CK-MB

CK-MB活性(U/L)(U/L)
activity
CK-MB

105

100

Residual activity (U/L)
CK-MM
CK-MM残存活性(U/L)

製造後安定性
Shelflife stability
CK-M阻害率(%)
Inhibition rate (%)
CK-M

CK-MM残存活性(U/L) (U/L)
Residual activity
CK-MM

阻害能比較
Comparison

CK-MB活性(U/L)
activity(U/L)
CK-MBSample : Human purified CK-MM

アスコルビン酸
Ascorbic acid
140120100806001020304050

アスコルビン酸 濃度(mg/100ml)
Concentration(mg/100mL)

Kanto Diagnostics
MI Judgement flow due to the value of CK, CK-MBJudgement flow
CK ≧ Limit of reference range

Yes

NoNo damage
Re-examine after 4 hours in the case of seizure of MI

CK-MB ≧ 25 IU/L

Measure LD if 2 -3 days passed since seizure of MINo

Yes

Skeletal muscle damage
Re-examine after 4 hours in the case of seizure of MI

6% ≦ CK-MB / CK ≦ 25%YesNoCK-MB活性(U/L)
CK-MB (U/L)

Myocardialdamage

CK-MB / CK ＜ 6%

Skeletal muscle damage

CK-MB / CK ＞ 25%

Macro CK, CK-BB

Guide of MI Judgement（*）

400① CK-MB/CK＜6％Skeletal muscle damage

CK-MB/CK > 25%
non-MI
Macro CK orCK-BB

300③②② 6％≦CK-MB/CK ≦25％
MI suspected

MI suspected

200③ CK-MB/CK＞25％
Macro CK or CK-BB

①100

25

CK-MB/CK < 6%
non-MI
Skeletal muscle damage

0010002000
3000
Limit of reference range
CK (U/L)
CK活性(U/L)*) Stein W：Laboratory diagnosis of acute
myocardial infarction, 30-154, E.Merck,
GIT Verlag, Darmstadt, 1988

4000

Product list
CODE
PRODUCT NAME
77494
CicaLiquid CK-MB (HMR)
77495
77798
CicaLiquid CK-MB(EPS)
77799
77822
CicaLiquid CK-MB(7170)
77823

R1
R2
R1
R2
R1
R2

PACKAGE
45mL×315mL×370mL×270mL×135mL×210mL×2CODEPRODUCT NAME

77826

CicaLiquid CK-MB(SP)77959

CicaLiquid CK-MB(008)

Storage：2 - 8℃

Expiry：8 months

R1R2
R1R2PACKAGE
20mL×27mL×261mL×322.5mL×3