▌常見Q&A

Q:關於如何選擇抗體方案?

A:

1. 貴實驗室目前要進行的是單株抗體製備or多株抗體製備?

    請詳見連結->單株抗體※Peptide單株抗體:from Peptide to mAb, 4~5 months
             ※蛋白質單株抗體:from Protein to mAb, 4~5 months

2. 目前已有antigen可進行多株抗體製備(客戶自行提供antigen)

     Antigen: protein->詳見※蛋白質多株抗體製備:from Protein to Ab, ~2 months

     Antigen: Plasmid->詳見※蛋白質表現純化多株抗體製備:from Expression& Purification Protein to Ab, 4~5 months

3. 須請曜鴻合成antige(曜鴻提供antigen合成服務)

     1peptide->請詳見※1peptide+多株抗體製備:from 1 Peptide to Ab,

                                    ※1peptide+多株抗體製備+親和性管柱純化: from 1 Peptide to Ab,      

     2peptide->請詳見※2peptides+多株抗體製備:from 2Peptides to Ab,

     Fusion protein->請詳見※重組蛋白多株抗體製備:from Fusion Protein to Ab, 4~5 months    

 

    (或者歡迎來電洽詢曜鴻02-26686845,立即有服務專員建議您方案選擇)

Q:關於Antigenicity prediction

A:Some peptide sequences are antigenic, some are not.  It is difficult to predict with 100% accuracy, because anitgenicity is a very complicate issue.  However many theories have been developed and many software programs are available to predict antigenicity of peptide sequences. 

1. Antigenicity prediction.  If you have a peptide sequence for antibody production, we can run a program to predict its antigenicity. We have used this program for many years to suggest sequences to use as antigens to produce antibodies.  It has been very useful.  Our customers like it very much.  We not just only find sequences, but also review it from the point of hyhobicity, aggregation, steric hindrance, etc.  The purpose is to ensure the quality of peptides as well as antibodies.   

(Note: Although our prediction of antigenicity has been very successful for many peptides and our customers are satisfied for the antiserum produced, the prediction of antigenicity is only a suggestion.  It is not a guarantee.)

2. Hyhobicity and hyhilicity.

3. Special conjugation and others. 

Q:關於曜鴻抗原(Peptide品質數據)

A:

※HPLC分析數據                                                                                      ※MASS分析數據

Q:關於Conjugation to a protein carrier

A:

OVA

Ovalbumin is the main protein found in egg white, making up 60-65% of the total protein. Ovalbumin displays sequence and three-dimensional homology to the serpin superfamily, but unlike most serpins it is not a serine protease inhibitor. The function of ovalbumin is unknown, although it is presumed to be a storage protein.

The ovalbumin protein of chickens is made up of 385 amino acids, and its relative molecular mass is 45 kDa. It is a glycoprotein with 4 sites of glycosylation. It is secreted from the cell. Unlike the signal sequences of most secreted proteins, Ovalbumins signal sequence is not cleaved. Additionally it is not located directly on the N-terminus, but rather internally, starting at residue 21 and ending at residue 47.

 

KLH

Keyhole limpet hemocyanin (KLH) is an extremely large, heterogeneous glycosylated protein consisting of subunits with a molecular weight of 350,000 and 390,000 in aggregates with molecular weights of 4,500,000-13,000,000. multisubunit, oxygen-carrying, metalloprotein found in the hemolymph of the giant keyhole limpet, Megathura crenulata, that lives off the coast of California from Monterey Bay to Isla Asuncion off Baja California.

Keyhole limpet hemocyanin (KLH) is used extensively as a carrier protein in the production of antibodies for research, biotechnology and therapeutic applications. Haptens are substances with a low molecular weight such as peptides, small proteins and drug molecules that are generally not immunogenic and require the aid of a carrier protein to stimulate a response from the immune system in the form of antibody production.[3] KLH is the most widely employed carrier proteins for this purpose. KLH is an effective carrier protein for several reasons. Its large size and numerous epitopes generate a substantial immune response, and abundance of lysine residues for coupling haptens, allows a high hapten:carrier protein ratio increasing the likelihood of generating hapten-specific antibodies. In addition, because KLH is derived from the limpet, a gastropod, it is phylogenetically distant from mammalian proteins, thus reducing false positives in immunologically based research techniques in mammalian model organisms.

 

BSA

Bovine serum albumin (BSA) is one of the most commonly used carriers for conjugation in antibody production. It belongs to the class of serum proteins called albumins, which make up about half of the protein in plasma and are the most stable and soluble proteins in plasma. It is very popular with laboratories developing immunoassays, mostly due to its availability, solubility and the numerous functional groups present for coupling to heptans.

    Molecular weight (MW): The MW of BSA is 67 X 103 which is much smaller than that of KLH, another commonly used carrier protein. It is, however, much more soluble and is immunogenic. It contains 59 lysines, and 30-35 of these are primary amines capable of reacting with a conjugation sites of linkers. It is a popular carrier for weakly antigenic compounds.

Solubility in aqueous solution: BSA is easily dissolved in an aqueous solution. It can be highly purified to become homogeneous. BSA is commonly used as protein standard in protein assays and as a molecular weight standard for SDS-PAGE and gel permeation chromatography.

 BSA can be used to block nonspecific binding sites in many immunochemical experiments such as ELISA, immunoblotting and immunohistochemical studies. It may be used as a non-relevant protein in enzyme immunoassays KLH cannot be used for this because the anti-KLH antibodies, which formed during immunization, will interfere with the measurement of anti-heptan antibodies. When KLH is used as the carrier, heptan-BSA conjugates can be used because they do not interfere with the measurement for anti-heptan antibodies.

Q:關於曜鴻抗體品質數據

A:

Western Blot Assay

Dot Blot Assay

Q:關於曜鴻近期抗體製備成功案例

A:

100.02   Anti-peptide抗體訂製(rabbit+抗體) 長庚大學測試結果抗體效果優秀!

100.01   Anti-peptide抗體訂製(rabbit+抗體) 屏東科技大學免疫結果經過測試後,客戶滿意!

99.12     Anti-peptide抗體訂製(peptide+ rabbit抗體) 中興大學成功!

99.11     Anti-peptide抗體訂製(peptide+ rabbit抗體) 中央研究院測試結果優秀!

99.09     Anti-peptide抗體訂製(peptide+ rabbit抗體) 海洋大學測試結果表現佳!

99.08     抗體訂製服務(rabbit+抗體) 國防醫學院測試結果表現極佳!

99.08     Anti-peptide抗體訂製(rabbit+抗體) 屏東科技大學客戶滿意度百分百!

99.01     多株抗體訂製服務 (rabbit +抗體) 淡江大學測試結果表現佳!!

99.03     抗體訂製服務(rabbit抗體) 國防醫學院測試結果令客戶滿意!!

99.04     Peptide抗體訂製(peptide+ guinea pig抗體) 海洋大學成功辨識原生蛋白!!

99.04     Peptdie抗體訂製(peptide+rabbit抗體) 海洋大學測試結果表現佳!!

99.05     Anti-peptide抗體訂製(peptide+ rabbit抗體) 中央研究院客戶測試結果滿意度佳!!

99.05     多株抗體訂製服務 (rabbit +抗體) 中央研究院測試結果優秀!!

99.06     Anti-peptide抗體訂製(peptide+ rabbit抗體) 中央研究院測試結果表現佳!!

99.07     Anti-peptide抗體訂製(peptide+ rabbit抗體) 中興大學成功辨識原生蛋白!!

99.08     Anti-peptide抗體訂製(rabbit+抗體) 屏東科技大學免疫結果經過測試後,客戶滿意度百分百!!

99.09     多株抗體訂製(表現純化+ rabbit抗體) 中央研究院測試結果表現佳!!

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