High Zone Tolerization Method

Creative Biolabs now can provide subtractive immunization service for our customers all over the world. One of the most important steps in the protocol of subtractive immunization is tolerization. In general, there are three tolerization methods: high zone tolerization, neonatal tolerization, and drug-induced tolerization. Now we are a focus on the high zone tolerization for the specific conditions.

Introduction of High Zone Tolerization

The method of high zone tolerization takes good advantages when the animals are injected with large quantities of soluble, non-aggregated antigens without the presence of adjuvants. The main mechanism is to prevent the formation of plasma cells which can produce antibody via interference with T-cells or B-cells as they two are all necessary for the activation of humoral immune response.

In general, the protocol of high zone tolerization can be organized into four steps:

(1) The immature repertoire of B-cells develops unique antigen specificities.
(2) After the introduction of large quantities of soluble, non-aggregated antigens without adjuvants, the corresponding T-cells cannot provide the secondary stimulus so that prevent the activation of the corresponding B-cells.
(3) Following the tolerization, an immunogen which contains a mixture of antigens including the tolerogen is introduced.
(4) The antigenically minor epitope is targeted via the immune response as there is no immune response for the immunodominant epitope.

High Zone Tolerance (Zijlstra, A.; et al. 2003).Fig.1 High Zone Tolerance (Zijlstra, A.; et al. 2003).

Applications and Limitations of High Zone Tolerization

According to high zone tolerization, one can direct the immune response to the desired antigen but avoid the complex manipulation of the host animals. What’ more, this tolerization method can strongly shorten the subsequent immunization time of the animals. In recent years, high zone tolerance has been extensively studied. On one hand, this method has been used to generate the specific antibodies via manipulating the immune response. On the other hand, it can even be used by some viruses for persisting in the host cells. However, there are also some limitations to this tolerization method. Firstly, large quantities of the tolerogen must be available in the form of pure, soluble, and non-aggregated. Secondly, the molecular composition of the tolerogen and immunogen should be simple. Thirdly, when the desired antigen is a minor component in the complex mixture, this method will be largely limited. In these conditions, we need to choose the other tolerization methods.

Creative Biolabs is a leading service provider that focuses on subtractive immunization. Now we can offer the high zone tolerization method for the specific immunization purposes. Based on our extensive experience and advanced platform, we can design and perform the custom protocol for your projects. If you are interested in our service, please do not hesitate to inquire us for more details.


  1. Zijlstra, A.; et al. Targeting the proteome/epitome, implementation of subtractive immunization. Biochemical & Biophysical Research Communications. 2003, 303(3): 733-44.

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