Our water-soluble QuickAntibody series of immunological adjuvants are divided into four types: 5-week immunological adjuvant for mice, 3-week immunological adjuvant for mice, 2-week immunological adjuvant for mice, and 8-week immunological adjuvant for rabbits. The 5-week adjuvant for mice has the best antibody affinity and antibody titer. Therefore, when preparing monoclonal antibodies, we generally recommend using the 5-week adjuvant for mouse immunization. For the preparation of polyclonal antibodies in mice, we generally recommend using a 3-week immunological adjuvant for mice. If time is tight or the quality requirements for antibodies are not high, mouse monoclonal antibodies can be prepared using 3-week mouse immunological adjuvants, and mouse polyclonal antibodies can be prepared using 2-week mouse immunological adjuvants.

Compared with the conventionally used Frejens adjuvant, QuickAntibody immunoadjuvant has multiple advantages such as a short immune cycle, fewer injections, no need for emulsification, low antigen dosage, high antibody titer, high antibody affinity, and easy acquisition of conformational epitope antibodies. The details are as follows: QuickAntibody only requires two doses of immunization, whether for the preparation of monoclonal or polyclonal antibodies, and can reduce the number of immunization doses compared with Frejner's adjuvant. 2.QuickAntibody can significantly reduce the total amount of antigen used by decreasing the number of immunization doses and the amount of antigen per dose. The recommended dosage of the antigen is as follows: (1) For subunit protein antigens with relatively weak immunogenicity, the dosage is 5-50 per dose. g (usually 5-20 μ g for mice and 20-50 μ g for rabbits); (2) Inactivated whole virus or whole bacteria and virus-like particle antigens with strong immunogenicity, 1-10 μ m per injection; g (usually 1-5 g for mice and 5-10 g for rabbits). 3.QuickAntibody features rapid antibody production, high antibody titer and strong antibody affinity. Take the standard immunization program as an example. Whether it is for the preparation of monoclonal or polyclonal antibodies, only two doses of immunization are needed within three weeks. Usually, a high affinity antibody level with an ELISA titer (Cutoff value of 0.1000) as high as 1:10,000-1:10,000,000 can be obtained by the fifth week. 4.QuickAntibody does not disrupt the natural conformation of the antigen, thus making it easy to screen and obtain monoclonal antibodies targeting conformational antigenic epitopes. This is an important feature that Frexner's adjuvant does not possess. 5.QuickAntibody is a water-soluble adjuvant. When in use, it does not require the complex emulsification process of Freund's adjuvant. The antigen and adjuvant only need to be simply mixed to immunize animals. 6.QuickAntibody uses the muscle immune pathway, which greatly facilitates its application compared with the conventional mouse monoclonal antibody preparation process that uses foot pads or endospleen immunity. One important use of QuickAntibody is that it can be conveniently used to prepare polyclonal antibodies for mice. Conventional polyclonal antibody preparation mostly uses rabbits. Not only are there many immunization shots, large amounts of antigen used and slow antibody production, but also due to the high technical requirements, it is often necessary to entrust specialized units to prepare them. With the QuickAntibody adjuvant, any laboratory animal personnel can conveniently and quickly prepare mouse polyclonal antibodies. The standard immunization program only requires simply immunizing 5 mice, and 1ml of high-quality mouse polyclonal antibodies can be obtained after five weeks. It is strongly recommended to use this adjuvant to prepare mouse polyclonal antibodies. 1ml of high-quality mouse polyclonal antibodies is sufficient to meet the experimental needs of the vast majority of users, including for enzyme-linked immunosorbent assay (ELISA), Western blotting, flow cytometry, immunohistochemistry and immunoprecipitation, etc.

小鼠的免疫，我们一般建议是50ul佐剂+50ul抗原，也可以适当增加，或左右腿各一针，这样共计是100ul佐剂+100ul抗原；也可以适当降低一些佐剂的用量，佐剂里面的有效成分是富余的，根据情况可以降低20-30%一般问题不大。但是不建议抗原用量过大，比如说直接100ug以上去免疫小鼠，抗原不会得到充分利用，动物反应比较强烈，甚至死亡。

每次免疫使用佐剂量为50ul，共两次免疫，因此1ml大概可以免疫8只小鼠 标准流程：1只小鼠免疫两针，50ul佐剂+50ul抗原，共100ul免疫1针，所以一只小鼠的佐剂用量为100ul。我们佐剂1ml一管，理论可以免疫10只，但是因为使用注射器损坏，离心管粘附等影响。只能免疫大概8只左右小鼠。

现混现用、拿到动物房混匀，混完后10min内免疫到动物体内，时间一长，沉淀会缓慢下沉，有效成分分布不均，会对免疫效果造成影响；客户也可以在实验室先预混，到动物房后直接用枪头吹打混匀或上下摇晃混匀，不能用注射器吹打混匀，混不开。 如何免疫：博奥龙特色QuickAntibody水佐剂，免疫小鼠的部位是后腿根部内侧肌肉，是单点注射免疫，小鼠腿部肌肉不太多，针刺入不要太深，容易刺刀骨头； 冲击免疫方法：腹腔注射抗原20~50ug,体积在100~200ul。腹腔注射点在小鼠左侧或右侧的下腹部，避开肝脏和较低中腹部膀胱，沿腿的角度穿透腹膜。1ml注射器针头进入1/3左右，有刺空感。

我们佐剂有成分是后续用于人疫苗佐剂的，所有优化都是使用肌肉免疫优化，因此建议使用肌肉免疫，但是也可以使用皮下一点或二点免疫，不建议腹腔免疫。

我们的佐剂是小鼠后退肌肉免疫，因部分客户之前使用腹腔免疫或皮下多点免疫，做肌肉免疫生疏，会鼓包，鼓包主要原因是打到了皮下，客户可以再扎深一些。初次使用，建议用酒精棉先擦拭后腿，免疫时避开后腿内侧两个静脉血管，可以在免疫后，将针头旋转半圈，静置几秒，拔出

一般不建议缩短，因为涉及到抗体的亲和力与效价问题，但是测血清效价，我们说明书是在2免后14天，时间上可以在2免后7-10天测效价，效价达标后几款冲击免疫做融合

质粒不行，蛋白和病毒可以

我们的佐剂经过数万种抗原免疫实验，目前的抗原种类有细胞、病毒、蛋白、多肽、多糖、脂类、重金属、抗生素、农药、毒品、食品安全类化合物等的免疫，效果均很好，有丰富的工业客户案例以及数百篇文献支持。

核酸免疫，我们佐剂不适用，可以采用脂质体

我们上架的佐剂现在仅供科研用途的，但是佐剂成分开发的最初目的的确是用于人用疫苗佐剂，是有成分是用于疫苗研发的。考虑到客户做动物痉苗的市场成本要求，一般很少客户考虑原料较为昂贵的佐剂，除非用于宠物与特种动物。如您有合作意向，可以联系我们进一步沟通咨询。

沉淀是我们佐剂的有效成分，不能够离心去除，去年的佐剂沉淀较少，今年佐剂沉淀较多，佐剂在21年11月份做了更新换代，新的佐剂以及验证5年以上时间，效果更好。

可以提供一个英文的原理描述：QuickAntibody is a proprietary novel adjuvant designed specifically for the preparation of monoclonal as well as polyclonal antibodies in mice. It is formulated with PAMPs (pathogen associated molecular patterns) and cationic polymers to activate cytosolic PRRs (pathogen recognition receptors).

一般免疫原性较弱的抗原或初次使用我们产品的客户，2针后效价可能起不来，此时可以在2免后14天进行第三针免疫，一般可以有一个数量级的增长，但是最多免疫4针，4针后效价仍未起来，一般建议重新免疫或修改抗原。比较常见的是小分子抗原如多肽和小分子。多肽一般可以建议aa数量增加到20个以上，可以有效的增加抗体效价。

这个正常，传统的佐剂含有卡介苗，卡介苗是强免疫原，脾脏中很多B细胞是针对卡介苗的，所以就会大一些

小鼠五周佐剂是最先出来的，在小鼠五周佐剂的基础上优化出来3周佐剂与2周佐剂，主要成分一致，配比略有不同。因此一般不建议混用。兔子与小鼠佐剂差异大

我们佐剂成分非常稳定，做过50℃一个月测试，无影响。但是不能冻存，冻存对免疫效果会有较大影响。一般保存可以放在4℃冰箱靠门的一次，不放在最里面，部分冰箱温控不好，放到最里面温度低于0℃.

For our rabbit polyclonal antibody project, there are two purification methods: (1) Antigen affinity purification Generally, purifying 10ml of antiserum can yield 1 to 3mg of the purified antibody. One rabbit can take 50ml of antiserum, so the total amount should be around 5 to 15mg. (2) ProiIenA/G purification Purifying 20ml of antiserum should yield approximately 100 to 150mg of purified antibodies. Generally, more customers adopt antigen affinity purification, and the previous data is mainly used. (ps: These are the results of Frexil adjuvant immunity.)

可以的，不同佐剂的功能是类似的，不同佐剂可以尝试交叉使用

可以结合使用，任何免疫方式都可以产生细胞免疫与体液免疫的效果。

The 1-10ug in the instruction manual refers to the total protein quantification of all bacteria or viruses. However, the total protein may be impure and very mixed, and it is uncertain whether it can effectively produce antibodies against the target protein. If it is pure protein, 1 microgram or even several hundred ng is sufficient

So far, QuickAntibody has been used to immunize various species such as mice, rats, rabbits, guinea pigs, goats, alpacas, chickens, and monkeys, and the results have all been good. The adjuvant itself is a mixture of multiple adjuvants rather than a single adjuvant. Species Adjuvant Type Antigen dosage ug Antigen Volume ul Adjuvant Volume ul Mouse QuickAntibody-Mouse 5-40ug 50 50 Rat, Guinea Pig QuickAntibody-Mouse 20-50ug 70 70 Rabbit, Chicken QuickAntibody-Rabbit 50-100ug 100 100 Goat, alpaca QuickAntibody-Rabbit 200-500ug 300-500 300-500 Monkey QuickAntibody-Rabbit 200ug 200-300 200-300

For the immunization of mice, we generally recommend 50 μ l of adjuvant +50 μ l of antigen. It can also be appropriately increased, or one injection can be given to each leg, so that the total is 100 μ l of adjuvant +100 μ l of antigen. The dosage of the adjuvant can also be appropriately reduced. The effective components in the adjuvant are in excess. Generally, reducing it by 20-30% is not a big problem depending on the situation. However, it is not recommended to use too much antigen. For instance, if more than 100 micrograms are directly used to immunize mice, the antigen will not be fully utilized, and the animals will have a strong reaction, and may even die.

说明书里的1-10ug是指的是全细菌或全病毒的总蛋白定量，但是总蛋白可能是不纯，非常杂，是否有效产生目标蛋白抗体，不确定。如果是纯的蛋白，1ug，甚至几百ng就够了

目前QuickAntibody免疫过：小鼠、大鼠、兔子、豚鼠、山羊、羊驼、鸡、猴子等多种物种，效果均不错。佐剂本身是多种佐剂的混合物，而不是单一佐剂。

The amount of adjuvant used for each immunization is 50 μ l, and there are two immunizations in total. Therefore, 1ml can immunize approximately 8 mice. Standard procedure: One mouse is immunized with two injections, 50 μ l of adjuvant and 50 μ l of antigen, totaling 100 μ l for one injection. Therefore, the adjuvant dosage for one mouse is 100 μ l. Each 1ml tube of our adjuvant can theoretically immunize 10 animals, but due to the damage of the syringe used and the adhesion of the centrifuge tube, etc., it has been affected. Only about 8 mice can be immunized.

Mix and use immediately. Take it to the animal room and mix it evenly. Immunize it into the animal's body within 10 minutes after mixing. If left for a long time, the precipitate will slowly sink, and the distribution of effective components will be uneven, which will affect the immunization effect. Customers can also premix in the laboratory first. After entering the animal room, they can directly mix by pipette tip or shake up and down to mix. Do not use a syringe to mix by pipette, as it will not mix well. How to immunize: With the special QuickAntibody water adjuvant of Boao Long, the site for immunizing mice is the inner muscle of the hind leg root. It is a single-point injection immunization. The leg muscles of mice are not too many, so the needle should not be inserted too deeply to avoid piercing the bone. Shock immunoassay: Intraperitoneal injection of 20 to 50 micrograms of antigen, with a volume of 100 to 200 micrograms. The intraperitoneal injection site was located in the lower abdomen on the left or right side of the mouse, avoiding the liver and the lower and middle abdominal bladder, and penetrated the peritoneum along the Angle of the leg. When the 1ml syringe needle enters about one-third, there is a feeling of being pierced through. "

Some of our adjuvants are used as adjuvants for human vaccines later on. All optimizations are carried out using muscle immunization. Therefore, it is recommended to use muscle immunization, but subcutaneous one-point or two-point immunization can also be used. Intraperitoneal immunization and tail vein immunization are not recommended.

Our adjuvant is for mouse hind leg muscle immunization. Due to some clients' previous use of intraperitoneal immunization or subcutaneous multi-point immunization, they are not familiar with muscle immunization and may experience bulges. The main reason for the bulges is that the injection has reached the subcutaneous layer. Clients can apply it deeper. For the first use, it is recommended to wipe the hind leg with an alcohol cotton swab. During immunization, avoid the two veins on the inner side of the hind leg. After immunization, rotate the needle half a circle, let it stand for a few seconds, and then pull it out.

It is generally not recommended to shorten the time as it involves the affinity and titer of antibodies. However, for the measurement of serum titer, our instructions state that it should be done 14 days after the second immunization. In terms of time, the titer can be measured 7 to 10 days after the second immunization. Once the titer reaches the standard, several impulse immunological tests can be combined

Plasmids won't work, but proteins and viruses can

Our adjuvants have been tested through tens of thousands of antigen immunization experiments. Currently, the types of antigens available for immunization include cells, viruses, proteins, polypeptides, polysaccharides, lipids, heavy metals, antibiotics, pesticides, drugs, and food safety compounds, all of which have achieved excellent results. We have a wealth of industrial customer cases and hundreds of literature articles to support this.

For nucleic acid immunization, our adjuvant is not applicable. Liposomes can be used instead.

Some of our adjuvant components are used for vaccine research and development, but the cooperation will not be completed until the second half of 2022. Our adjuvant components are used for the development of human vaccine adjuvants. Generally, customers cannot afford to use them for animal vaccines, except for pets and special animals.

The precipitate is the effective component of our adjuvant and cannot be removed by centrifugation. Last year, there was less precipitate in the adjuvant, but this year there is more. The adjuvant was updated and replaced in November 2021. The new adjuvant has been verified for more than five years and has better effects.

This adjuvant is a confidential formula of our company. The specific ingredients are not available for disclosure. An English description of the principle can be provided QuickAntibody is a proprietary novel adjuvant designed specifically for the preparation of monoclonal as well as polyclonal antibodies in mice. It is formulated with PAMPs (pathogen associated molecular patterns) and cationic polymers to activate cytosolic PRRs (pathogen recognition receptors)."

For general antigens with weak immunogenicity or for customers using our product for the first time, the titer may not increase after two doses. At this point, a third dose can be administered 14 days after the second dose, which usually leads to an increase of an order of magnitude. However, the maximum number of doses is four. If the titer still does not increase after four doses, it is generally recommended to re-administer or modify the antigen. The more common ones are small molecule antigens such as polypeptides and small molecules. It is generally recommended to increase the number of aa peptides to more than 20, which can effectively enhance the antibody titer.

This is normal. Traditional adjuvants contain BCG vaccine. BCG vaccine is a strong immunogen. Many B cells in the spleen target BCG vaccine, so they are larger

The five-week adjuvant for mice was the first to come out. Based on the five-week adjuvant for mice, the three-week adjuvant and the two-week adjuvant were optimized. Their main components were the same, but the ratios were slightly different. Therefore, it is generally not recommended to mix them. There are significant differences in adjuvants between rabbits and mice Mouse adjuvants and rabbit adjuvants are not recommended to be mixed. One of the main components of adjuvants has species selectivity. Mouse adjuvants themselves can be mixed, but if there is enough time, it is not recommended to mix them.

Just looking at the price, it's a bit expensive, but it saves time cost Save one month's time cost and the cost of raising mice 2. Save the amount of antigen used, only 1/5 to 1/10 of the original. The price of 1mg of natural antigen can reach tens of thousands of yuan. Our adjuvant components are very stable. We have conducted a one-month test at 50℃ and found no effect. However, it cannot be frozen, as freezing can have a significant impact on the immune effect. For general storage, it can be placed once near the door of a 4℃ refrigerator, not at the innermost part. Some refrigerators have poor temperature control, and placing it at the innermost part will keep the temperature below 0℃. Sure. The functions of different adjuvants are similar, and different adjuvants can be tried to be used alternately

Our adjuvant components are very stable. We have conducted a one-month test at 50℃ and found no effect. However, it cannot be frozen, as freezing can have a significant impact on the immune effect. For general storage, it can be placed once near the door of a 4℃ refrigerator, not at the innermost part. Some refrigerators have poor temperature control, and placing it at the innermost part will keep the temperature below 0℃.

我们这边兔多抗项目，有两种纯化方式： （1）抗原亲和纯化 一般纯化10ml抗血清能获得1~3mg纯化后抗体，一只兔子能取50ml抗血清，这样总量应该在5~15mg左右； （2）ProiIenA/G纯化 纯化20ml抗血清应该能拿到100~150mg左右纯化后抗体。 一般客户采用抗原亲和纯化的多一些，以前一个数据为主。（ps:这些是弗式佐剂免疫的结果）

Sure. The functions of different adjuvants are similar, and different adjuvants can be tried to be used alternately

They can be used in combination. Any immune method can produce the effects of cellular immunity and humoral immunity.

The main function of impulse immunity is to activate spleen cells to a relatively high extent, which has a significant impact on the ability of hybridoma cells to secrete antibodies after fusion. Therefore, direct antigen impulse immunity is still recommended. Moreover, cells themselves as immunogens are likely to have the problem of low titer, and their components are rather diverse. The target epitope protein may not necessarily be the dominant immunogen. Previously, General Manager Yan said that taking on similar technical services for preparing antibodies through cellular immunity basically could not meet the experimental expectations.

There is no weight requirement. Currently, when our company immunizes rabbits, we use healthy 6-week-old New Zealand white rabbits (with bright eyes and clean butts, weighing about 2 kilograms).

目前QuickAntibody免疫过：小鼠、大鼠、兔子、豚鼠、山羊、羊驼、鸡、猴子等多种物种，效果均不错。佐剂本身是多种佐剂的混合物，而不是单一佐剂。

物种	佐剂类型	抗原用量ug	抗原体积ul	佐剂体积ul
小鼠	QuickAntibody-Mouse	5~40ug	50	50
大鼠、豚鼠	QuickAntibody-Mouse	20~50ug	70	70
兔子、鸡	QuickAntibody-Rabbit	50~100ug	100	100
山羊、羊驼	QuickAntibody-Rabbit	200~500ug	300~500	300~500
猴子	QuickAntibody-Rabbit	200ug	200~300	200~300

Glycoprotein immunogens, such as CA125, are highly prone to generating IgM and difficult to obtain IgG

The surface of bacteria is all polysaccharides, making it difficult to obtain protein epitope antibodies. Polysaccharide epitope antibodies have a high titer. It can be attempted to immunize 1-5 micrograms after quantification by BCA

Rabbit adjuvants and mouse adjuvants should not be mixed. They are species-specific and it is not recommended to use them together