Predicting the adsorption of proteins by aluminium-containing adjuvants

Seeber, Sally J.; White, Joe L.; Hem, Stanley L.

doi:10.1016/0264-410x(91)90154-x

Cited by 132 publications

(75 citation statements)

References 6 publications

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“…The adsorption of proteins to aluminum hydroxide and aluminum phosphate adjuvants is a complex phenomenon, depending on the surface charge of the protein and adjuvant as well as other factors [24][25][26][27]. In this case, charge did not appear to be the major factor driving adherence, as greater than 90% adherence could be achieved at pH 5.5-6.5, under which conditions both the protein and adjuvant had net positive charge.…”

Section: Discussionmentioning

confidence: 99%

Improved formulation of a recombinant ricin A-chain vaccine increases its stability and effective antigenicity

Carra

Wannemacher

Tammariello

et al. 2007

Vaccine

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Section: Discussionmentioning

confidence: 99%

Improved formulation of a recombinant ricin A-chain vaccine increases its stability and effective antigenicity

Carra

Wannemacher

Tammariello

et al. 2007

Vaccine

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“…It has been established that optimal binding of protein to Al(OH) 3 occurs when the pH of the diluent matrix is between the isoelectric point of the protein in question and the point of zero charge of the aluminum salt (11)(12)(13). Because the isoelectric point of rAls3p-N was similar to the pH levels of saline and PBS, it is not surprising that the levels of protein binding were similar in both diluents.…”

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confidence: 94%

Considerable Differences in Vaccine Immunogenicities and Efficacies Related to the Diluent Used for Aluminum Hydroxide Adjuvant

Lin

Ibrahim

Avanesian

et al. 2008

Clin Vaccine Immunol

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We are developing an anticandidal vaccine using the recombinant N terminus of Als3p (rAls3p-N). We report that although more rAls3p-N was bound by aluminum hydroxide diluted in saline than by aluminum hydroxide diluted in phosphate-buffered saline (PBS), its immunogenicity and efficacy were superior in PBS. Thus, protein binding, by itself, may not predict the efficacy of some vaccines with aluminum adjuvants.Hundreds of millions of doses of aluminum adjuvants have been administered to humans over the past century (5,11,12). One standard formulation is Alhydrogel, a gelatinous matrix of aluminum hydroxide, which minimizes lot-to-lot variability and enables facile adsorption of proteins (5, 12). As noted by Hem et al., aluminum hydroxide in its dehydrogenated, crystalline form is chemically aluminum oxyhydroxide [AlO(OH)] (7), but in its aqueous phase, it acquires an additional water molecule to become aluminum trihydroxide [Al(OH) 3 ].We have developed a vaccine using the recombinant N terminus of the candidal adhesin Als3p (rAls3p-N), which has been shown to protect mice against otherwise-lethal disseminated candidiasis (8,9,15,16). The vaccine is also effective in combination with Alhydrogel diluted in phosphate-buffered saline (PBS) (8). Vaccine efficacy requires an adjuvant, as the vaccine does not significantly improve survival if no adjuvant is utilized. To determine the optimal diluent for the continued development of the rAls3p-N vaccine, we assessed its relative binding to Al(OH) 3 , its immunogenicity, and its efficacy in the presence of PBS versus in the presence of saline. rAls3p-N (amino acids 17 to 432 of Als3p) was produced in Saccharomyces cerevisiae and purified by Ni-nitrilotriacetic acid matrix affinity purification as previously described (15). The isoelectric point of several batches of rAls3p-N was determined to be 7.1 using a Ready Gel IEF pH 5-to-8 gel (Bio-Rad) per the manufacturer's instructions (data not shown).To quantify protein binding to Al(OH) 3 , rAls3p-N was diluted to 0.2 mg/ml in saline (pH 6.5, 0.9% sodium chloride) or PBS (pH 7.40, 0.9% sodium chloride, 0.1% sodium-potassium phosphate), with or without 0.13 mg/ml of Alhydrogel (Brenntag Biosector, Frederikssund, Denmark). The solutions were vortexed, incubated for 30 min at room temperature, and centrifuged at 13,000 rpm for 10 min on a tabletop microcentrifuge. The protein concentration in the supernatant (i.e., unbound) was calculated by UV absorption as follows: (concentration without Alhydrogel -concentration with Alhydrogel)/(concentration without Alhydrogel). Protein binding was found to be slightly but significantly higher in the presence of saline than in the presence of PBS (median

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“…Protein adsorption occurs by several mechanisms, including hydrogen bonding, hydrophobic bonding, electrostatic bonding and van der Waals' force. Seeber et al (13) concluded that electrostatic forces have a major role in protein adsorption of aluminum hydroxide and that an extensive binding between antigen and adjuvant was seen when they were oppositely charged. Under the present conditions, Al-and Ca-gels may have been charged positively since OVA (pI = 4.6) was charged negatively.…”

Section: Discussionmentioning

confidence: 99%

Relationship between Hemolytic Activity and Adsorption Capacity of Aluminum Hydroxide and Calcium Phosphate as Immunological Adjuvants for Biologicals

Katô

Shibano

Saito

et al. 1994

Microbiology and Immunology

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Aluminum hydroxide (Al) and calcium phosphate (Ca) gels have been used as vaccine adjuvants for many years. We investigated mechanism of the hemolytic activities of both adjuvant materials. The hemolytic activity of each gel depended on the gel dose. The adsorption capacities and the hemolytic activities of both adjuvants decreased as the concentration of phosphate increased in a gel-washing solution. A positive correlation between the hemolytic activity and the adsorption capacity was found in Al-gel. A disruptive effect of Ca-gel on membrane of erythrocytes was shown by electron microscopy. Ca-gel required more than 10 times as much pre-adsorbed ovalbumin as did Al-gel to inhibit the hemolysis. These results suggest that the hemolytic activity of both adjuvant materials depended mainly on the adsorption ability, and it may be useful to control the adsorption ability of adjuvants to reduce their hemolytic activity.

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Predicting the adsorption of proteins by aluminium-containing adjuvants

Cited by 132 publications

References 6 publications

Improved formulation of a recombinant ricin A-chain vaccine increases its stability and effective antigenicity

Improved formulation of a recombinant ricin A-chain vaccine increases its stability and effective antigenicity

Considerable Differences in Vaccine Immunogenicities and Efficacies Related to the Diluent Used for Aluminum Hydroxide Adjuvant

Relationship between Hemolytic Activity and Adsorption Capacity of Aluminum Hydroxide and Calcium Phosphate as Immunological Adjuvants for Biologicals

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