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.

Celiac Disease Versus Gluten Sensitivity – New Role For Genetic Testing and Fecal Antibody Testing?

Celiac disease (CD) has a prevalence of 1/100. Between 90-99% of Celiacs are HLA DQ2 and/or DQ8 positive. Every individual has two DQ serotypes. Because the molecular HLA nomenclature can be confusing DQ serotyping is a method for simplifying the results. There are four major types and 5 subtypes: HLA DQ1, DQ2, DQ3 and DQ4; DQ1 has two subtypes; DQ5 and DQ6 whereas DQ3 has three subtypes; DQ7, DQ8 and DQ9. Each individual has two copies of HLA DQ. One DQ type is inherited from each parent.

Though 35-45% of individuals of Northern European ancestry are DQ2 &/or DQ8 positive only 1% have classic CD as defined by abnormal blood tests and small intestine biopsies. Several autoimmune conditions also occur more frequently in DQ2 and DQ8 positive individuals.

There is accumulating scientific evidence that many individuals are gluten sensitive and respond to a gluten free diet though they have normal blood tests and/or normal intestinal biopsies (fail to meet strict criteria for CD). This is more commonly being referred to as non-Celiac gluten sensitivity (NCGS). Many individuals who have NCGS are relatives of confirmed Celiacs and were previously referred to as latent Celiacs. Electron microscopy and immunohistochemistry studies of individuals with normal biopsies but suspected of or at risk (1st degree relatives of Celiacs) have revealed ultrastructural abnormalities of the intestine and those who chose a gluten free diet usually responded and many who did not ultimately developed abnormal biopsies on long term follow-up. Seronegative Celiac has also been recognized, that is blood tests are negative, but the biopsy reveals classic abnormalities of Celiac and the individual responds to gluten free diet.

Testing for DQ2/DQ8 has been suggested as a way to exclude CD. That is, if you are negative for DQ2 and DQ8, then you are very unlikely to have CD. However, well documented cases of CD and Dermatitis Herpetiformis (DH) have been confirmed in DQ2 and DQ8 negative individuals. Moreover, we now have the clinical experience that other DQ patterns predispose a person to gluten sensitivity because these individuals frequently have elevated fecal antibodies to AG or tTG and respond to a gluten free diet.

Why some people develop Celiac Disease or become gluten sensitive is not well understood. Risk factors include onset of puberty, pregnancy, stress, trauma or injury, surgery, viral or bacterial infections including those of the gut, medication induced gut injury or toxicity (e.g. NSAIDs), immune suppression or autoimmune diseases, and antibiotic use resulting in altered gut flora (dysbiosis). The severity of the sensitivity is related to the DQ type, pre-existing intestinal injury, degree of exposure to gluten (how frequent and large a gluten load an individual is exposed to), and immune status. Once initiated, gluten sensitivity tends to be lifelong. True CD requires lifelong complete gluten avoidance to prevent serious complications, cancers, and early death.

Serotypes can be determined from blood or buccal mucosal cells (obtained by oral swab) from several commercial labs including Prometheus, Labcorp, Quest, The Laboratories at Bonfils, and Enterolabs. Fecal IgA anti-gliadin and IgA tissue transglutaminase antibody testing is only available in the U.S. commercially through Enterolabs. The fecal AG and tTG testing may be helpful in those with normal blood tests for Celiac and/or a normal small bowel biopsy but suspected of being gluten sensitive. Though the fecal antibody results are not widely accepted by many “Celiac experts” numerous testimonials of individuals testing positive only on fecal tests who have responded to gluten free diet can be found in support groups, web postings, personal communication from Dr. Fine and this physician’s clinical experience.

Fecal antibody testing for gliadin (AG) and tissue transglutaminase (tTG) by Enterolab in Dallas has revealed elevations in 100% of Celiacs tested and up to 60% of symptomatic individuals without Celiac disease (NCGS) even if not DQ2 or DQ8 positive. The only DQ pattern he found not associated with gluten sensitivity is DQ4/DQ4, a pattern typically found in non-Caucasians who are known to have a low prevalence of Celiac disease.


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Type 2 Diabetes – Can Low Levels of Antibodies Contribute to the Development of Diabetes?

Scientists at Guandong Medical University and several other research institutions in China and Scotland found low levels of certain types of antibodies in people who had been diagnosed with Type 2 diabetes. These antibodies stop inflammation, which has a known link to insulin resistance, the cause of Type 2 diabetes.

In November of 2017, the Journal of Inflammation, (London), reported on a study that looked at antibodies to inflammatory molecules in people with Type 2 diabetes. Women had decreased levels of the antibodies called anti-IL6 IgG and anti-IL8 IgG. Both molecules are inflammatory, as is anti-TNF-ALPHA IgG, which was seen in low levels in men. The participants were then given blood sugar lowering drugs for 6 months. HbA1c levels were found to be…

  • lowest in those diabetics with the highest levels of anti-ILALPHA IgG and
  • highest in those with low levels of the antibodies,

although the differences were not statistically significant.

From this information the researchers concluded deficiencies of antibodies to inflammatory molecules probably raises the risk for the development of Type 2 diabetes.

Antibodies are proteins that attach themselves to molecules known as antigens. We usually think of antibodies as connecting themselves to alien organisms, such as bacteria and viruses to enable white blood cells to engulf and kill the invaders.

Inflammation is part of the immune response, but it must fade away after the invaders have been vanquished. The role of antibodies to inflammatory molecules is to stop inflammation.

Antibodies are also known as immunoglobulins, and they are composed of three protein chains held together in a Y shape. Five types of antibody are classified according to their species of chains. The chains include…

  • IgG,
  • IgM,
  • IgA,
  • IgE, and
  • IgD.

Inflammation is defined by the five conditions it causes…

  • heat,
  • redness,
  • pain,
  • loss of function, and
  • swelling.

Inflammation in the body is named for the inflamed organ plus the ending “itis.” For instance, inflammation of the skin is called dermatitis, “derma” for skin, and “itis” for inflammation.

Two main classes of drugs are used for fighting inflammation…

  • cortisone which resembles a body hormone, and
  • non-steroidal anti-inflammatory drugs (NSAIDs) include medications such as aspirin, Tylenol, and Motrin.

Cortisone is a steroid, which works against inflammatory reactions by blocking white blood cells from going to the site where the inflammation takes place. (Not to be confused with steroids used illegally to boost athletic performance).

NSAIDs work against body molecules called prostaglandins, which are also pro-inflammatory.

The Real Reasons Doctors Don’t Check Antibody Levels in Hypothyroid Patients

Logic dictates that if the most common cause of hypothyroidism is from an autoimmune mechanism, then the first test ran on all newly diagnosed hypothyroid patients would be a thyroid antibody test. Well logic doesn’t always reign supreme and in many cases Hashimoto’s Autoimmune thyroid patients go undiagnosed and just given the standard thyroid replacement treatment. I am going to explore the most likely reasons this occurs.

The first and foremost reason most thyroid patients will never have their antibody levels checked is because the treatment protocol does not change based on the mechanism. Most doctors are going to give you synthetic thyroid replacement hormones, intermittently check your TSH, and call it a day. If you continue to suffer with the same symptoms as before they will be attributed to some other ailment, usually depression or anxiety, and be treated accordingly.

The second reason for the failure to order antibody tests, is a branch off of the first reason. Since the treatment is not going to be dictated based off the antibody results (although it should), then running the test becomes “medical waste” in the eyes of the insurance companies. The payment for a test that will not alter the course of treatment is in a way wasteful, but the real waste is in ignoring the underlying cause of the problem and leaving the patient to suffer. Many times patients are mocked and scorned for suggesting a test or treatment that is a bit outside of the tight little box Hashimoto’s patients are placed in.

Having no alternative treatment plan for Autoimmune thyroid, and the resultant medical waste of a test ordered but that has no bearing on the patients treatment are the two main reasons many Hashimoto’s patients go undiagnosed and ignored in the system.

Determining that hypothyroidism is from an immune mechanism, IS important, and should absolutely be managed in more comprehensive way, not only including some type of replacement hormone, but it should also include management of the immune systems destruction of the gland itself. A more comprehensive approach gives the patient a better chance to feel and function normally.