ELISAELISA
1
2
Ag Ab Ag-Ab Ag AB:
Ka [Ag-AB] [Ag][Ab][ g ] [ g][ ]
Kd Ka
Ka M-1
Kd M
KdKd
3
Kd: R
HH
4
100
5
ELISAELISA
ELISA
6
Kd 10-15M
7
Biotin 244.3.
B H
Coenzyme R
lysine -lysine (biocytin)
8
Avidin68 00068,000
4 128 N4 128 NC
Avidin
11
Kd 10-15M
9
ELISA
10
11
12
RIA
RIA
CV 2~3CV 2 3
13
ELISA
2
14
RIA ELISA
STD
0.1 0.063 0.1004 0.06426 0.1047 4.28
0.25 0.105 0.2496 0.1071 0.2572 3.04
0.5 0.199 0.5018 2.034 0.5124 2.11
1 0.426 0.9977 0.4345 1.0159 1.82
2.5 1.173 2.5119 1.1965 2.5612 1.96
5 2.206 4.9683 2.25 5.0902 2.49
10 3.558 10.0479 3.629 10.4286 3.79
15
16
KdKd
17
ELISA
1000010000
ELISA
18
Std. 1 Std. 2 Std. 3 Std. 4 Std. 5 Std. 6 Std. 7 Std. 8 Std. 9
S lidSolidAb Kd 10pM 20 40 80 160 320 640 1280 2560
9.901 19.802 39.60 79.20 158.39 316.73 633.24 1265.51 2526.210000 100 99.0% 99.0 99.0 99.0 99.0 99.0 98.9 98.9 98.7
10000 3009.708
97.1%19.416
97.138.8397.1
77.6597.1
155.2797.0
310.3997.0
620.1696.9
1237.6396.7
2462.0296.210000 300 97.1% 97.1 97.1 97.1 97.0 97.0 96.9 96.7 96.2
10000 10009.090
90.9%18.179
90.936.35
90.972.68
90.9145.26
90.8290.12
90.7578.59
90.51150.05
89.82266.86
88.5
10000 30007.691
76.9%15.379
76.930.75
76.961.45
76.8122.73
76.7244.74
76.5486.56
76.0961.04
75.11869.97
73.0
4 999 9 995 19 98 39 92 79 68 158 72 314 88 619 54 1198 4110000 10000
4.99950.0%
9.99550.0
19.9850.0
39.9249.9
79.6849.8
158.7249.6
314.8849.2
619.5448.4
1198.4146.8
19
20
Kd 1000pM
21
22
5000pM 10000pM
23
24
25
ELISAELISA
26
ELISAELISA
27
ng/ml
ng/ml ng/ml (%) (%) ng/mlg g
10 0.9672 9.7 90.3 9.035 0.8273 16.55 83.45 4.17
2.5 0.588 23.52 76.48 1.9122.51.25 0.4844 38.75 61.25 0.7656
0.625 0.2868 45.89 54.11 0.33810 313 0.1444 46.21 53.79 0.16810.3130.156 0.086 55.13 44.87 0.07
28
50ng/ml10,000pM
10,000pMKd 1000pM
29
0.1244Ln(x)+0.075
30
31
32
2
33
122
T10000pM 1000pM 0.156 10ng/ml
27~1724pM10ng/ml pM
34
35
Edge Edgeg g
36
Blank absorbance obtained by 2operators
m0(
620)nm
Abs.
450
Well pos ition A 1 to H1
S tandard curves obtained by 2 operators
nm50
(62
0)Abs
45
Insulin concentration, pg/ml37
ELISA
38
IgG F(ab’)2, Fab’
Fc heavy chain
F(ab’)22 Fab’
39
Washing repetition and blank absorbance(Mean & SD)(Mean & SD)
) nm
450(
620)
banc
e Ab
srob
Assay kit: Mouse insulin ELISA KIT (U-type)Difference of mean absorbance: p<0.065 Difference of variance:p<0.1
40
Assay kit: Rat TSH ELISA KITDifference of mean absorbance: p<0.03 Difference of variance: p<0.25p p
41
Biotin-avidin
D galactosidase-D-galactosidase4-methylumbelliferyl- -D-galactoside2 it h l D l t id2-nitrophenyl- -D-galactoside
Horseradish peroxidaseo se ad s pe o dase3-(4-hydroxyphenyl)propionic acid
42
ELISA RIAELISA RIA
pM
pM
43
44
45
ELISA
46
90 l
No. C
10 l
No. CNo. C
47
No. C ng/ml
x0.9 0.1 ng/ml
48
49
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