Biomedical subjects

Anandi Krishnan

Publications and source records attributed to Anandi Krishnan.

8 recordsLinked to original sources

PubMed2006

Interfacial energetics of globular-blood protein adsorption to a hydrophobic interface from aqueous-buffer solution.

Adsorption isotherms of nine globular proteins with molecular weight (MW) spanning 10-1000 kDa confirm that interfacial energetics of protein adsorption to a hydrophobic solid/aqueous-buffer (solid-liquid, SL) interface are not fundamentally different than adsorption to the water-air (liquid-vapour, LV) interface. Adsorption dynamics dampen to a steady-state (equilibrium) within a 1 h observation time and protein adsorption appears to be reversible, following expectations of Gibbs' adsorption isotherm. Adsorption isotherms constructed from concentration-dependent advancing contact angles theta(a) of buffered-protein solutions on methyl-terminated, self-assembled monolayer surfaces show that maximum advancing spreading pressure, Pi(a)max, falls within a relatively narrow 10 < Pi(a)max < 20 mN m(-1) band characteristic of all proteins studied, mirroring results obtained at the LV surface. Furthermore, Pi(a) isotherms exhibited a 'Traube-rule-like' progression in MW similar to the ordering observed at the LV surface wherein molar concentrations required to reach a specified spreading pressure Pi(a) decreased with increasing MW. Finally, neither Gibbs' surface excess quantities [Gamma(sl)-Gamma(sv)] nor Gamma(lv) varied significantly with protein MW. The ratio {[Gamma(sl)-Gamma(sv)]/Gamma(lv)} approximately 1, implying both that Gamma(sv) approximately 0 and chemical activity of protein at SL and LV surfaces was identical. These results are collectively interpreted to mean that water controls protein adsorption to hydrophobic surfaces and that the mechanism of protein adsorption can be understood from this perspective for a diverse set of proteins with very different composition.

Adsorption↗

PubMed2006

Interfacial rheology of blood proteins adsorbed to the aqueous-buffer/air interface.

Concentration-dependent, interfacial-shear rheology and interfacial tension of albumin, IgG, fibrinogen, and IgM adsorbed to the aqueous-buffer/air surface is interpreted in terms of a single viscoelastic layer for albumin but multi-layers for the larger proteins. Two-dimensional (2D) storage and loss moduli G(') and G(''), respectively, rise and fall as a function of bulk-solution concentration, signaling formation of a network of interacting protein molecules at the surface with viscoelastic properties. Over the same concentration range, interfacial spreading pressure Pi(LV) identical with gamma(lv)(o)-gamma(lv) rises to a sustained maximum Pi(LV)(max). Mixing as little as 25 w/v% albumin into IgG at fixed total protein concentration substantially reduces peak G('), strongly suggesting that albumin acts as rheological modifier by intercalating with adsorbed IgG molecules. By contrast to purified-protein solutions, serially diluted human blood serum shows no resolvable concentration-dependent G(')and G('').

Air↗

PubMed2006

Interfacial energetics of blood plasma and serum adsorption to a hydrophobic self-assembled monolayer surface.

Interfacial energetics of blood plasma and serum adsorption to a hydrophobic, methyl-terminated self-assembled monolayer (SAM) surface (solid-liquid SL interface) are shown to be essentially the same as to the buffer-air interface (liquid-vapor LV interface). Specifically, spreading pressure (Pia) isotherms scaled on a w/v concentration basis constructed from advancing contact angles (thetaa) of serially diluted plasma/serum derived from four different mammalian species (bovine, equine, human, and ovine) on the SAM surface are not resolvable at the 99% confidence level and furthermore are found to be strikingly similar to isotherms of purified human-blood proteins. Maximum advancing spreading pressures Piamax for protein mixtures fall within a relatively narrow 17<Piamax<26 mN/m band, mirroring results obtained at the LV surface. These observations lead to the conclusion that neither depletion of coagulation proteins in the conversion of plasma to serum nor variation in the plasma proteome among species has a substantial affect on adsorption energetics to these test hydrophobic surfaces. Experimental results are rationalized on the basis that there is a generic mechanism controlling adsorption of globular-blood proteins to test hydrophobic surfaces. We conclude that this generic mechanism is the hydrophobic effect by which proteins are expelled from aqueous solution in order to increase hydrogen-bonding (self-association) among water molecules at the expense of less favorable water-protein interactions. Expelled protein readily displaces water within the hydrophobic-surface region and becomes adsorbed. The amount of water displaced per gram of adsorbed protein does not vary greatly among globular proteins because the partial specific volume v0 of globular proteins is quite conserved (0.70<or=v0<or=0.75 cm3/g protein). Any single blood protein or mixture of proteins consequently displaces nearly an equivalent amount of interfacial water and hence adsorption is observed to scale similarly with solution concentration expressed in w/v units.

Adsorption↗

PubMed2005

Scaled interfacial activity of proteins at a hydrophobic solid/aqueous-buffer interface.

Contact-angle goniometry confirms that interfacial energetics of protein adsorption to the hydrophobic solid/aqueous-buffer (solid-liquid, SL) surface is not fundamentally different than adsorption to the aqueous-buffer/air (liquid-vapor, LV) interface measured by pendant-drop tensiometry. Adsorption isotherms of 9 globular blood proteins with molecular weight (MW) spanning from 10 to 1000 kDa on methyl-terminated self-assembled monolayer surfaces demonstrate that (i) proteins are weak surfactants, reducing contact angles by no more than about 15 degrees at maximum solution concentrations ( approximately 10 mg/mL); (ii) the corresponding dynamic range of spreading pressure Pi(a) < 20 mN/m; and (iii) the maximum spreading pressure Pi(max) (a) for these diverse proteins falls within a relatively narrow 5 mN/m band. As with adsorption to the LV interface, we find that concentration scaling substantially alters perception of protein interfacial activity measured by Pi(a). Proteins appear more similar than dissimilar on a weight/volume basis whereas molarity scaling reveals a systematic ordering by MW, suggesting that adsorption is substantially driven by solution concentration rather than diversity in protein amphilicity. Scaling as a ratio-to-physiological-concentration demonstrates that certain proteins exhibit Pi(max)(a) at-and-well-below physiological concentration whereas others require substantially higher solution concentration to attain Pi(max)(a). Important among this latter category of proteins is blood factor XII, assumed by the classical biochemical mechanism of plasma coagulation to be highly surface active, even in the presence of overwhelming concentrations of other blood constituents such as albumin and immunoglobulin that are shown by this work to be among the class of highly surface-active proteins at physiologic concentration. The overarching interpretation of this work is that water plays a dominant, controlling role in the adsorption of globular-blood proteins to hydrophobic surfaces and that energetics of hydration control the amount of protein adsorbed to poorly water-wettable biomaterials.

Adsorption↗

PubMed2005

An evaluation of methods for contact angle measurement.

A systematic comparison of Wilhelmy-balance tensiometry (WBT), tilting-plate goniometry (TPG) and captive-drop goniometry (CDG), using a variety of silane-treated glass surfaces of variable wetting characteristics is reported. WBT was assumed to be the benchmark of comparison (gold standard). Advancing angles measured by TPG and CDG were in statistical agreement with corresponding WBT measurements. In contrast, receding angles from both goniometric techniques were systematically higher than WBT.

Chemistry, Physical↗

PubMed2005

Liquid-vapor interfacial tension of blood plasma, serum and purified protein constituents thereof.

A systematic study of water-air (liquid-vapor, LV) interfacial tension gamma(lv) of blood plasma and serum derived from four different mammalian species (human, bovine, ovine and equine) reveals nearly identical concentration-dependence (dgamma(lv)/dlnC(B); where C(B) is plasma/serum dilution expressed in v/v concentration units). Comparison of results to a previously-published survey of purified human-blood proteins further reveals that dgamma(lv)/dlnC(B) of plasma and serum is surprisingly similar to that of purified protein constituents. It is thus concluded that any combination of blood-protein constituents will be substantially similar because dgamma(lv)/dlnC(B) of individual proteins are very similar. Experimental results are further interpreted in terms of a recently-developed theory emphasizing the controlling role of water in protein adsorption. Accordingly, the LV interphase saturates with protein adsorbed from bulk solution at a fixed weight-volume concentration ( approximately 436 mg/mL) independent of protein identity or mixture. As a direct consequence, dgamma(lv)/dlnC(B) of purified proteins closely resembles that of mixed solutions and does not depend on the relative proportions of individual proteins comprising a mixture. Thus variations in the plasma proteome between species are not reflected in dgamma(lv)/dlnC(B) nor is serum different from plasma in this regard, despite being depleted of coagulation proteins (e.g. fibrinogen). A comparison of pendant-drop and Wilhelmy-balance tensiometry as tools for assessing protein gamma(lv) shows that measurement conditions employed in the typical Wilhelmy plate approach fails to achieve the steady-state adsorption state that is accessible to pendant-drop tensiometry.

Animals↗

PubMed2004

Mixology of protein solutions and the Vroman effect.

Mixing rules stipulating both concentration and distribution of proteins adsorbed to the liquid-vapor (LV) interphase from multicomponent aqueous solutions are derived from a relatively straightforward protein-adsorption model. Accordingly, proteins compete for space within an interphase separating bulk-vapor and bulk-solution phases on a weight, not molar, concentration basis. This results in an equilibrium weight-fraction distribution within the interphase that is identical to bulk solution. However, the absolute interphase concentration of any particular protein adsorbing from an m-component solution is 1/mth that adsorbed from a pure, single-component solution of that protein due to competition with m - 1 constituents. Applied to adsorption from complex biological fluids such as blood plasma and serum, mixing rules suggest that there is no energetic reason to expect selective adsorption of any particular protein from the mixture. Thus, dilute members of the plasma proteome are overwhelmed at the hydrophobic LV surface by the 30 classical plasma proteins occupying the first 5 decades of physiological concentration. Mixing rules rationalize the experimental observations that (i) concentration-dependent liquid-vapor interfacial tension, gammalv, of blood plasma and serum (comprised of about 490 different proteins) cannot be confidently resolved, even though serum is substantially depleted of coagulable proteins (e.g., fibrinogen), and (ii) gammalv of plasma is startlingly similar to that of purified protein constituents. Adsorption-kinetics studies of human albumin (66.3 kDa) and IgM (1000 kDa) binary mixtures revealed that relatively sluggish IgM molecules displace faster-moving albumin molecules adsorbing to the LV surface. This Vroman-effect-like process leads to an equilibrium gammalv reflecting the linear combination of weight/volume concentrations at the surface predicted by theory. Thus, the Vroman effect is interpreted as a natural outcome of protein reorganization to achieve an equilibrium interphase composition dictated by a firm set of mixing rules.

Adsorption↗

PubMed2004

Scaled interfacial activity of proteins at the liquid-vapor interface.

A principal conclusion drawn from observations of time- and concentration-dependent liquid-vapor (LV) interfacial tension gamma(lv) of a diverse selection of proteins ranging from albumin to ubiquitin spanning nearly three decades in molecular weight (MW) is that concentration scaling substantially alters perception of protein interfacial activity as measured by reduction in gamma(lv). Proteins appear more similar than dissimilar on a weight/volume basis, whereas molarity scaling reveals a "Traube-rule" ordering by MW, suggesting that adsorption is substantially driven by solution concentration rather than diversity in protein amphilicity. Scaling as a ratio-to-physiological-concentration demonstrates that certain proteins exhibit the full possible range of interfacial activity at and well-below physiological concentration, whereas others are only weakly surface active within this range, requiring substantially higher solution concentration to achieve reduction in gamma(lv). Important among this latter category of proteins are the blood factors XII and XIIa, assumed by the classical biochemical mechanism of plasma coagulation to adsorb to procoagulant surfaces, even in the presence of overwhelming concentrations of other blood constituents such as albumin and immunoglobulin that are shown by this work to be among the class of highly surface-active proteins at physiologic concentration. A comparison of pendant drop and Wilhelmy balance tensiometry as tools for assessing protein interfacial activity shows that measurement conditions employed in the typical Wilhelmy plate approach fails to achieve the steady-state adsorption condition that is accessible to pendant drop tensiometry.

Adsorption↗