Yes, however, Doxorubicin itself is a fluorescent molecule therefore you must be careful not to use a fluorescent tag that is within the same range (excitation wavelength 480 nm; emission wavelength 580 nm). Most of the regular colored fluorescent tags are not compatible. You need to make sure that your fluorescent tag will not have any excitation or emission that will interfere with Doxorubicin.
Liposomes are particles so even empty liposomes without the drug are going to cause some level of immune reaction. At least for one of your experiments you need to use control plain liposomes. Also if you are using Immunodox products you need to use the regular Doxorubicin liposomes as control as well. Surface modified Doxorubicin liposomes and regular Doxorubicin liposomes have different bio-distribution in body.
The yield of conjugation is different using different reactive lipids. Usually maleimide lipids have higher yield of conjugation with antibody and antibody fragments, however the molecule should have available reactive sulfur. Some conjugation chemistries are multi-step and complex and some are based on simple incubation. The conjugation can be through the N-terminus or C-terminus of antibody or antibody fragments. The conjugation can be through the available sulfur on Fab’ fragments or thiolated antibodies. The conjugation can be done through the sugar molecules on the antibody or through click chemistry. Each conjugation chemistry serves its own purpose. Before ordering the kits you need to do some research about the type of your antibody as well as your target site.
In some instances the best way to determine the most effective conjugation method is through trial and error. You may also be able to refer to our database for helpful suggestions.
This is not necessarily true. Myocet which is a Non-PEGylated Doxorubicin liposome has shown in clinical trials remarkable tumor shrinkage capability. Myocet is commercially available drug that is mainly marketed in Europe. PEGylated Doxorubicin is also effective and is marketed as Doxil in the United States and Lipodox in Asia.
Non-PEGylated products require the remote loading of the active ingredient, Doxorubicin, using a pH gradient. The pH gradient allows the maximum amount of Doxorubicin to be loaded into the plain Doxosome or Immunodox liposomes. One drawback of this loading technique is the weak stability of the pH gradient. Therefore immediately before drug administration you will use the 3 vials in the kit to create a fresh pH gradient.
In addition, Non-Pegylated liposomes are created with unsaturated phospholipids which form loosely packed liposomes around the encapsulated drug. This means that once the drug is loaded into the liposome it slowly leaks its contests in a matter of a week inside the vial. By using the 3 vial kit and loading the Doxorubicin directly before administration all of the encapsulated drug will be delivered to the test subjects.
The PEGylated products on the other hand use a ammonium sulfate gradient that remains stable over time. They are also formed using saturated phospholipids which form a much more compact liposome around the encapsulated drug. Therefore the liposomes can be pre-loaded with Doxorubicin.
There are two different types of Immunodox kits. Immunodox-NP (Non-PEGylated) which comes in three vial kits and Doxorubicin drug should be loaded to the liposomes right before the liposomes are administered to the animal. The kits come with eleven different reactive lipids that can be used for various types of chemistries to antibodies and antibody fragments.
PEGylated Immunodox formulation are made from saturated lipids and Doxorubicin drug is already loaded into the liposomes. The PEGylated Immunodox come with 12 different reactive lipids that can be used for various types of chemistries to antibodies and antibody fragments.