Custom Antibody Sequencing – Discovery of the Century

Brief introduction to Custom antibodies

Antibodies help us to protect from numerous bacterial infections. They are big protein compounds white cells. Custom antibodies are distinctive proteins which are separated from the analysis subject’s blood when a particular disorder or foreign compound is inserted into its structure. Initially, custom antibodies were described in studies carried by experts in early 1980s. To accelerate to express Charles Darwin’s concept of evolution, editors of New Scientist Magazine, John Gribbin and Jeremy Cherfas, introduced an impressive technique to research the human development. They produced a blood serum protein and inserted it into a rabbit. The entire body of the rabbit identified the foreign proteins and created antibodies – which were particular to unique proteins. You will find many companies which develop custom antibodies and provide high-end services like transient expression or antibody sequencing for scientists who are trying to understand and explore vaccines for various health issues. Recognized as custom antibody solutions, these agencies generally get samples of the foreign element and the subject’s serum or blood. The foreign element is recognized as an antigen and the antibodies are proteins which combine with them.

Antibody sequencing and transient expression at a glance

Many companies exist today which deals with protein sequencing and sequencing dead hybridomal cells. The researchers which want to boost their process of innovation and support, approaching such companies that deal with transient expression is the right option. Today, the antibody sequencing is performed over different species like rat, mouse, human guinea pig, hamster, bovine, rabbits and human antibody concealing EBV cell lines. Most of the companies which are into the antibody development business maintain the highest standards and follow projected timelines.

Transient expression of a transformed gene in the preferred target cells over the comparatively short time period; and doesn’t always show integration of a gene into the host chromosome and isn’t transferred to the following generation. For the fast and quick formation of cell lines indicating full-length recombinant antibodies and completely improved antibodies, modern science has introduced enhanced technology.

For the period of the transient expression, the protein is indicated just for a short time, that is perfect for confirmation of theory because the DNA which is released in the transfection procedure is generally not involved into the nuclear genome and is going to be diluted via degraded or mitosis. When the scientist has recognized the subjected protein of interest, the development of a balanced transfected cell line is the expected second step that needs the transfected gene to live in cells’ genome. This takes more time compared to transient transfections to get the stable transfection. The multi-simultaneous testing method uses numerous proprietary vector models for increasing the development of recombinant protein you need. After that, the Bioinformatics is performed for creating best suited proteins required for specific studies.

Following the same, there are many companies these days which deals with transient expression and antibody sequencing as well as helps in challenging projects like DNA sequencing, drug development, cloning, patent application, etc. Today, the advanced antibody development plays a vital role in helping human beings fight against insoluble disease and treat the patients.

Humanized Monoclonal Antibody Therapies in Neoplastic Disease

In the last ten years the treatment of many types of cancer has been revolutionized by monoclonal antibody therapies. There are many, many print and audiovisual advertisements for these drugs bringing them into the public mainstream.. So, this brings to point some important questions: 1) What are monoclonal antibodies? 2) What types of cancer are treatable by this therapy? 3) What is the basic mechanism of action, i.e., How do these drugs work?

Antibodies are the body’s natural immune defense against invading pathogens. They are produced by specific cells of the immune system termed B cells. When an infection of the body occurs, the immune system takes note. After a brief time, when the front line defenses of the immune system battle the infection, [termed the innate immune response- the first line of battle], the cellular part of the fight takes over [the adaptive response]. The adaptive immune response involves antigen presenting cells, T and B cells. Very briefly, what happens within your body when an infection occurs is this: Circulating “detectives” called macrophages and dendritic cells find the infection and literally eat it or eat cells infected [in the case of a viral infection]. The cells then “present” specific protein parts of the eaten pathogen [bacteria or virus–called antigens] to cells called T and B cells. This is a call to arms: this is literally a scream saying “time to kick butt” by the immune system. The T cells are activated by the B cells upon finding the presenting antigen. The B cells then decide that they can better fight by changing themselves into a plasma cell. This is where the action happens. The B cell changes into a protein pumping machine, the protein being antibodies. The antibodies then go find from the blood the pathogen that the original antigen presenting cells early in the infection encountered. When the antibodies find their target, they bind to it and mark it for death. Think of this as a ball with toilet plungers attached. The wooden handle is what tells the immune system, “this thing needs to die”.

This is the basic theory behind monoclonal cancer therapy. Except in the context of monoclonal antibody cancer treatment, the antibodies are produced to a single epitope (a particular protein that the immune system readily sees from a cancer cell). These cells are then taken into the laboratory, grown and stimulated to produce a single antibody that “sees” a cancer cell.

An example of monoclonal antibody treatment in cancer is Herceptin. Herceptin is a monoclonal antibody specific for an antigen called HER2. HER2 is a protein that is more prevalent on cancer cells of the breast than in normal cells. This protein is from a family of receptors that tells normal cells to grow. The drug then takes advantage of this and can specifically target cancer cells of the breast in order to kill them. This drug too has side effects of being NOT heart friendly.

Another good example of neoplastic cancer therapy is the drug termed Rituxan (Rituximab). Rituxan is a monoclonal antibody directed towards the antigen CD20 on circulating lymphocytes of the blood and is indicated for the treatment of non-Hodgkin’s lymphoma. It works by depleting the blood of cells that have over-produced cells that have CD20 on their cell surface. When the antibody binds, like Herceptin, the body sees those cells as foreign and kills them.

So, the question is now that we know what neoplastic antibody therapy is, how does this killing work?
The current research and thinking is that when cancer cells are coated by these antibody drugs, the extending end of the antibody [called the Fc portion] attracts Natural Killer, NK, cells from the immune system. These cells actually have receptors that recognize this very event. When NK cells find antibody coated cells, they bind to them tightly and commence to kill them. The close proximity of the two cells allows the NK cell to release protein degradating enzymes, and other cytotoxic elements to kill the target.
This entire process is termed Antibody Dependent Cellular Toxicity, ADCC.

ADCC is a powerful tool being utilized by numerous biotech companies with the intent to augment the immune response with other, novel drugs. Such drugs that come to mind are TLR agonists, chemotherapy drugs and gene transfer strategies.

Genentech makes Herceptin, while Rituximab is in combination with Biogen Idec/Genentech. Both drugs are multimillion dollar assets to both companies. The side effects of Herceptin have been documented and are under intense research to understand them. Rituxan continues to be a key element to the bottom line of both companies with it’s successful treatments.

Herceptin is produced by Genentech.
Rituxan is another monoclonal antibody treatment indicated for the treatment of lymphoma.

Antisperm Antibodies

There are many causes and reasons for infertility in males, of which one of them is antisperm antibodies. A man suffering from antisperm antibodies generally gets sensitized to sperm, to cause an immune system that destroys the sperms found in the body.

In a normal man, a barrier in his testes protects the sperm from the immune system. However, those men with antisperm antibodies have a broken barrier wherein immune cells can easily access the sperm. With the unique antigen surface that is found on such sperms, the immune system detects its presence and thus triggers a response.

About 10% of infertile men suffer from antisperm antibodies and this is what interferes with the quality and function of the sperm. It is depending on the location of the antibodies that the sperm is affected. If the antibodies are on the tail, then the sperms are either immobilized or found clumped together. If they are found on the head, then the sperm is prevented from binding to the egg, and thus prevents fertilization from taking place.

There are various causes for antisperm antibodies developing, of which the main reason is the cervical mucus found in some woman. This cervical mucus tends to develop antibodies in the sperm and is the cause of about 40% of the unexplained infertility in couples. Even anything that disrupts the barrier found between the sperm and immune system like infection, injury to the testicles and twisting of testicles can lead to antisperm antibodies.

Other causes of antisperm antibodies are undescended testicles, testicular biopsy and cancer, varicocele and congenital absence of vas deferens. About 70% of men who had a vasectomy reversal have also been found to have antisperm antibodies.

Doctors can easily diagnose antisperm antibodies with an analysis of the man’s sperm and by analyzing the woman’s cervical mucus after sex for any antibodies. However, when diagnosed with antisperm antibodies, it is not easy to eliminate the antibodies. High doses of cortico steroids generally lower the number of antibodies to restore fertility temporarily. However, these high doses bring with it many serious side effects.

This is the reason couples who suffer from this condition usually turn to assisted reproductive technologies to conceive a baby. One of the techniques used here is washing the sperm before in vitro fertilization. With this, the sperm can produce fertilized eggs better for implantation. Sometimes, washing of the sperm is also used in intrauterine insemination.

Paternal Leukocyte Immunization As Therapy for Excessive Sperm Antibodies

There are various factors preventing couples to have children, one of them is because excessive sperm antibodies of the wife. For these cases, PLI (Paternal Leukocyte Immunization) therapy could be an option to quickly have a baby.

PLI (Paternal Leukocyte Immunization) or also known as Husband Leukocyte Immunization Husband (HLI) is treatment to lower sperm antibodies in women who have excessive number of these antibodies.

Excessive sperm antibodies makes it difficult for sperm to get to the egg because it is always rejected and become dysfunctional, so it does not allow for fertilization and pregnancy. Every woman who has been exposed to sperm does have antibodies against the sperm of her husband, but in some women these antibodies overreact.

The cause is the same with people who are allergic, each person has a different response, depending on the individual. Women with high sperm antibodies overreact to proteins on the sperm, so sperm is rejected and become dysfunctional.

PLI therapy is given by injecting husband’s white blood cells to area under the skin of the wife. It aims to reduce the wife sperm antibodies that can be tolerated by the body and allow for fertilization.

The injection is given at least 3 times, each is given every 3 weeks. Serum containing husband’s white blood cells will be injected at the bottom of the mother’s skin. After therapy, patients are advised to do a reassessment of imuno-andrology test. If the result has reached the normal limit then it is not necessary to continue the treatment. If not, repeated therapy can be performed to achieve normal limits.

There are several requirements that must be considered by married couples to be able to perform PLI therapy:

  1. Excessive wife sperm antibodies
  2. Husband is in good health, i.e. a blood test showed that the husband is free from infectious diseases such as HIV, hepatitis and others.
  3. For the wife who has any history of allergy (e.g. seafood allergy), at the time of therapy, the allergy is not in relapsed condition. If therapy is being performed at the time of allergy it can cause an overreaction.

The chance of success in reduction of antibodies can be up to 95 percent, but for the chance of pregnancy depends on the condition of the body, respectively. Difficulty to get pregnant depends on many factors. If it is indeed because of high antibodies then there is high probability you can get pregnant after reduction of these antibodies.

Before PLI therapy was invented, there are 2 ways which are usually conducted to help pair with this problem to get pregnant, that is with the use of condoms and imunosupressor drug.

Women are expected to gradually lower her sperm antibodies by limiting the exposure to sperm by the use of condoms. However, this should take a long time, about 6 months to 1 year. This will be even longer if the wife often eat foods that contain protein allergens.

Another way is to use the imunosupressor drug. Unfortunately, by using this drug all the antibodies in the women’s body will also goes down, not only sperm antibodies. As a result, the wife will often experience illness, and even affect the spinal cord that causes slow blood rejuvenation, lead to osteoporosis, the risk of diabetes, and others.

It is later found that the husband’s white blood cell immunization can lower sperm antibodies exclusively, and keep the other antibodies which are needed. PLI makes good tolerance to sperm.

The wife who has this problem will also have difficulty in getting good result in IVF. This is because the antibodies will not only reject the sperm, but also reject the fetus resulted from fertilization of the sperm. Even if IVF yields successful conception, the percentage of miscarriage is still high. This is because the wife body remained resistant to the fetus.

Women with a history of this problem in the family had a greater risk of experiencing the same thing. A history of certain food or condition allergies also increases the risk of women having excessive sperm antibodies.