FastAb Adjuvant
Adjuvants and Antibody Production

FastAb Adjuvant is a proprietary, novel, water-soluble mouseadjuvant designed specifically for preparing monoclonal andpolyclonal antibodies. It can be used as an alternative to theclassic Freund’s adjuvant.
Compared to Freund’s adjuvant, FastAb Adjuvant utilizes
simpler preparation steps (thus shorter time), a lower dosageof immunogen, and no emulsification (and is hence capable ofmaintaining the native structure of the antigen).
FastAb Adjuvant can be applied to monoclonal Ab screeningand preparation with mouse hybridoma technology, phagedisplay antibody library technology or single B cell cloningtechnology.

SKU
08642901
Package
1ml / Vial
Storage
Stable for 24 months from the date ofmanufacture when stored at 2 - 8℃
(Do not freeze)
NoteThis product can be stored at ambient
temperature for shipping purposes
Applicable Species
Mouse

Highlight
1. Easy to use. No emulsification - just mix with antigen and it is ready to be used for intramuscular
injection.
2. Simple and short protocol. 2- or 3-weeks immunization with a total of 2 injections.
3. Reduced antigen usage, with a single dose of 0.1-50 µg.
4. Sterile, safe, and non-toxic; thus ensures good animal welfare.
5. Allows for the generation of antibodies with high titer and affinity.
6. As emulsification is not required, the native immunogen structure can be maintained. Suitable forboth linear and conformational epitopes.
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Use Method
1. For each mouse, prepare 0.1 - 50 µg of immunogen in 50 µL of normal saline.
Note: Please prepare sufficient solution according to the number of animals.
Recommended antigen usage:1) Strong immunogen: 0.1 - 10 µg2) Weak immunogen: 1 - 50 µg
2. Mix the FastAb Adjuvant thoroughly (by vortexing, if possible). Take the required volume of FastAb
Adjuvant and quickly mix the adjuvant and antigen-saline solution at a 1:1 ratio. The recommended
injection volume is 100 µL/mouse; prepare fresh mixture before the injection.
Note: It is normal to see some precipitates in FastAb Adjuvant and antigen-saline-FastAb Adjuvant
mixture. Ensure the mixture is well mixed before the injection.
3. The mixture shall be injected intramuscularly into the hind leg using a recommended volume of
100 µL/mouse.

Recommended Immunization Protocol
5-weeks standard protocol for generation of monoclonal antibody

Time PointDay 0Process
Primary immunization (single dose)Day 21

Booster immunization (single dose)Day 31-35

Bleed at day 10 - 14 after booster injection and measure the serum titer byELISA

Day 35

Pulse immunization without adjuvant

Day 36-38

Harvest spleen or lymph nodes from the mouse, screen and generate monoclonal
antibodies through cell fusion, followed by establishing the immune antibody
library or single B cell isolation

2-weeks standard protocol for generation of polyclonal antibody

Time Point
Day 0

Process
Primary immunization (single dose)

Day 7

Booster immunization (single dose)

Day 14

Collect the blood from caudal vein, eyeball or by cardiac puncture

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Performance Data
5-weeks standard protocol for generation of monoclonal antibody

Day 36 - 38

Day 35

Day 31 - 35
Day 21Day 0
Primaryimmunization(single dose)

Boosterimmunization(single dose)

Bleed at day 10 - 14
after booster
injection, measure
the serum titer byELISA

Pulse
immunization
without adjuvant

Harvest spleen orlymph nodes fromthe mouse, screen
and generatemonoclonal
antibodies throughcell fusion, followedby establishing theimmune antibody
library or single B cellisolation

2
1.8
1.6
1.4
1.2
1
0.8
0.6
0.4
0.2
0
1:
20
0
1:
40
0
1:
80
0
1:
1600
1:
32
00
1:
6400
1:
12
80
0
1:
25
60
0
1:
51
20
1:
10 0
24
0
1:
20 0
48
0
1:
40 0
96
0
1:
81 0
92
1:
16 00
38
40
pre-i
0mm
B
unle( ank
1:
1000
)OD

Antigen 1

Titer

Mouse #1

Mouse #2

Mouse #3

page 3 of 7

Mouse #4

Mouse #5

2
1.8
1.6
1.4
1.2
10.8
0.6
0.4
0.2
0
1:
20
0
1:
40
0
1:
80
0
1:
1600
1:
32
00
1:
64
00
1:
1280
0
1:
25
60
0
1:
51
20
1:
10 0
24
0
1:
20 0
48
0
1:
40 0
96
0
1:
81 0
92
1:
16 00
38
40
pre-i
0mm
B
unle( ank
1:1000)OD

Antigen 2

Titer

Mouse #1

Mouse #2

Mouse #3

Mouse #4

Mouse #5

2
1.8
1.6
1.4
1.2
10.8
0.6
0.4
0.20
1:
200
1:
40
0
1:
80
0
1:
1600
1:
3200
1:
6400
1:
1280
0
1:
25
60
0
1:
51
20
1:
10 0
240
1:
20 0
48
0
1:
40 0
96
0
1:
81 0
92
1:
16 00
38
40
pre-i
0mm
B
unle( ank
1:1000)OD

Antigen 3

Titer

Mouse #1

Mouse #2

Mouse #3

Mouse #4

Mouse #5

Fig 1. Determination of serum antibody titer (by ELISA) in the mice immunized with three different antigens (n=5 per antigen),i.e., Antigen 1 (B), Antigen 2 (C), Antigen 3 (D). Sera samples were collected at day 27; blank and sera collected at day 0 (preimmune) served as negative controls.

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8-weeks immunization protocol for weak immunogenDay 59Day 49Day 21

Day 0Primaryimmunization(single dose)

Boosterimmunization(single dose)

Day 42
Boosterimmunization(single dose)Day 56

Pulse
Bleed at dayimmunization
10 - 14 after
booster injection, without adjuvant
measure theserum titer by
ELISA

Harvest spleen or
lymph nodes fromthe mouse, screen
and generatemonoclonal
antibodies throughcell fusion, followedby establishing theimmune antibodylibrary or single B
cell isolation

2
1.8
1.6
1.4
1.2
1
0.8
0.6
0.4
0.2
0
1:
20
0
1:
40
0
1:
80
0
1:
1600
1:
32
00
1:
6400
1:
1280
0
1:
25
60
0
1:
51
20
1:
10 0
24
0
1:
20 0
48
0
1:
40 0
96
0
1:
81 0
92
1:
16 00
38
40
pre-i
0mm
B
unle( ank1:1000)

OD

Antigen 4

Titer

Mouse #1
Mouse #4

Mouse #2
Mouse #5

Mouse #3
Mouse #6

Fig 2. Determination of serum antibody titer (by ELISA) in the mice immunized with a weak immunogen, i.e., Antigen 4. Sera
samples were collected at day 49; blank and sera collected at day 0 (pre-immune) served as negative controls.

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Frequently Asked Questions
Question 01. What steps are recommended for achieving optimal performance prior to using
FastAb Adjuvant?
FastAb Adjuvant is a multi-component suspension; mix thoroughly before using. It isrecommended to inject as soon as possible after mixing the FastAb adjuvant with
antigen to avoid precipitation and ensure good performance.
Question 02. How do I determine immunogen usage?
1) Weak immunogen: 1 - 50 μg2) Strong immunogen: 0.1 - 10 μg
Question 03. When should the FastAb Adjuvant be used?
FastAb Adjuvant should be used for primary immunization and booster, and not for pulse
immunization.
Question 04. What types of antibodies do mice produce when using FastAb Adjuvant?
Using the 5-week standard immunization protocol, the antibodies produced by the mice
will be mainly high affinity IgG and some low affinity IgM.
Question 05. How do I design the immunization protocol for monoclonal antibodies when using
FastAb Adjuvant?
We recommend performing 5-week standard immunization protocol to generate themonoclonal antibody.
One extra immunization can also be added between day 35 - 42 based on the titer anduser’s requirement. If the titer measured at day 10 - 14 after the extra immunization
cannot meet the user’s requirement, it is suggested to redesign the antigen or increase
the number of mice and repeat the entire process from the beginning.
Question 06. Is it possible to use rapid immunization protocol when preparing mouse monoclonal
antibody with FastAb Adjuvant?
Weekly or biweekly injection may allow the immunized mice to produce higher level IgG.
However, this shortened immunization interval is not suitable for production of highaffinity antibodies; hence it is not recommended to use the rapid immunization protocol
for generating the mouse monoclonal antibodies.
Question 07. Is it possible to use rapid immunization protocol when preparing mouse polyclonal
antibody with FastAb Adjuvant?
As the serum volume in each mouse is small, a large-scale production of polyclonal
antibodies will not be feasible. If users only require a small amount of polyclonal
antibody, weekly or biweekly injection (with a total of two injections) can be performed.
Question 08. What kind of antibody screening or technology is suitable for preparing mousemonoclonal antibody with FastAb Adjuvant?
There are 3 options: mouse hybridoma technology, phage display antibody library
technology or single B cell cloning technology.

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Frequently Asked Questions
Question 09. Can FastAb Adjuvant be applied to prepare humanized antibodies?
Sure. FastAb Adjuvant can be applied to immunize the humanized mice initially, followedby use of mouse hybridoma tevchnology, phage display antibody library technology orsingle B cell cloning technology to prepare the humanized antibody.
Question 10. What are the main advantages of FastAb Adjuvant as compared to Freund’sadjuvant?
Ready-to-use, simple and easy preparation, with no emulsification and less immunogenrequirement.
Question 11. Is the immunogen for FastAb Adjuvant the same as that of Freund’s adjuvant?
Yes, both adjuvants can be used for various antigens.
Question 12. How is the performance of FastAb Adjuvant as compared to Freund’s adjuvant?
The performance is similar.
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