5. The prototype microporation system parameters are set to
create an array of micropores with the following details. Pore
density of 75 micropores/cm
2
, 80 μm wide, 300 μm long, and
40–50 μm in depth (see Note 5).
3.3.3 Vaccine Delivery
Using Shock Waves
1. Shock wave-based needle-free device is used as described
[2, 13].
2. Hair on the skin is removed in the abdomen region using a hair
clipper or commercial hair removal cream.
3. The vaccine to be delivered is loaded in the sample holder.
4. The delivery device is placed in a perpendicular position against
the shaved abdominal skin of the mice.
5. Shock wave formation is triggered, and liquid jet is accelerated
to high velocity.
6. Multiple shots are administered to increase the volume of
injection (see Note 6).
3.3.4 Vaccine Delivery
Using Microneedles [39]
1. Mice are anesthetized and their abdominal hair is closely
trimmed using an electric trimmer (no scraping of the skin
surface).
2. After trimming, the skin is wiped clean with alcohol and lightly
dried with a lint-free tissue. The mouse is then placed on the
animal interface plate with the abdomen facing down and is
positioned over an opening to allow access to the microporation tip assembly. A square array of micropores is then created
and is loaded with the drug to be injected (see Note 7).
4 Notes
1. Serum should be free of hemoglobin. Hemolysis should be
avoided for accurate results.
2. Fine gauge needle should be used to avoid collateral damage
and hostile immune response.
3. Appropriate care should be taken to protect the integrity of
DNA vaccine. It should not be vigorously pipetted/handled to
prevent shearing.
4. Spermidine treatment is extremely crucial for DNA binding to
gold nanoparticles. Improper incubation times might lead to
infective DNA-particle binding.
5. The pore size of microinjector determines the velocity and
depth of penetration of the vaccine. Pore size should be optimized according to the skin type and thickness for efficient
vaccine delivery.
444
Akshay Datey et al.
create an array of micropores with the following details. Pore
density of 75 micropores/cm
2
, 80 μm wide, 300 μm long, and
40–50 μm in depth (see Note 5).
3.3.3 Vaccine Delivery
Using Shock Waves
1. Shock wave-based needle-free device is used as described
[2, 13].
2. Hair on the skin is removed in the abdomen region using a hair
clipper or commercial hair removal cream.
3. The vaccine to be delivered is loaded in the sample holder.
4. The delivery device is placed in a perpendicular position against
the shaved abdominal skin of the mice.
5. Shock wave formation is triggered, and liquid jet is accelerated
to high velocity.
6. Multiple shots are administered to increase the volume of
injection (see Note 6).
3.3.4 Vaccine Delivery
Using Microneedles [39]
1. Mice are anesthetized and their abdominal hair is closely
trimmed using an electric trimmer (no scraping of the skin
surface).
2. After trimming, the skin is wiped clean with alcohol and lightly
dried with a lint-free tissue. The mouse is then placed on the
animal interface plate with the abdomen facing down and is
positioned over an opening to allow access to the microporation tip assembly. A square array of micropores is then created
and is loaded with the drug to be injected (see Note 7).
4 Notes
1. Serum should be free of hemoglobin. Hemolysis should be
avoided for accurate results.
2. Fine gauge needle should be used to avoid collateral damage
and hostile immune response.
3. Appropriate care should be taken to protect the integrity of
DNA vaccine. It should not be vigorously pipetted/handled to
prevent shearing.
4. Spermidine treatment is extremely crucial for DNA binding to
gold nanoparticles. Improper incubation times might lead to
infective DNA-particle binding.
5. The pore size of microinjector determines the velocity and
depth of penetration of the vaccine. Pore size should be optimized according to the skin type and thickness for efficient
vaccine delivery.
444
Akshay Datey et al.